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  • Original articles
    Mechanism of simulated microgravity based on RNA-seq technology
    QIN Lingmei, JIN Rui, ZHOU Ying, LI Jiangbo, LI Xinyue, CHENG Long
    2023, 47(7): 481-486. https://doi.org/10.7644/j.issn.1674-9960.2023.07.001
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    Objective To screen differentially expressed genes（DEGs） before and after simulated microgravity （SMG） using RNA sequencing （RNA-seq）, and to explore the mechanism of SMG for human cells at the transcriptomic level. Methods HL-60 cells were cultured under different culture conditions, the experiments were divided into three groups： the normal gravity（NG） group,simulated microgravity （SMG） group and SMG-NG（SMGNG） group. RNA-seq analysis was used to screen the differential expression profiles of mRNA in HL-60 cells before and after SMG.Gene ontology（GO） analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG）analysis were used for functional annotation to analyze the pathways regulated by SMG and those that could or could not recover rapidly after resumption of NG conditions. Results A total of 735 differentially expressed genes （DEGs） （424 up-regulated and 311 down-regulated） were identified in SMG comparing with NG, 624 of which rapidly recovered in SMGNG, while 110 DEGs failed to rapidly recover. GO and KEGG analysis found that DEGs that rapidly recovered were highly correlated with signal transduction, lipid metabolism, immune regulation and apoptosis, while those that did not rapidly recover were mainly involved in pathways related to nucleotide-binding oligomerization domain （NOD）-like receptor signal transduction, steroid metabolism and bone metabolism. Conclusion The target genes and signaling pathways regulated by SMG are identified, which will facilitate research on the biological functions and mechanisms of microgravity.
  • Original articles
    Construction and identification of a pancreatic specific Golgi protein 73 knockout mouse model based on Cre-LoxP system
    LI Jingfei, ZHANG Xiaojuan, XU Yixin, ZHANG Yanhong, WAN Luming, WEI Congwen, WANG Wei, ZHONG Hui
    2023, 47(7): 487-492. https://doi.org/10.7644/j.issn.1674-9960.2023.07.002
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    Objective To construct and identify a model of pancreatic-specific Golgi protein 73 （GP73） knockout mice for studies on GP73 function in the pancreas. Methods A model of GP73flox/+ mice was constructed by inserting two LoxP loci into both ends of exon 4 of the GP73 gene via homologous recombination in fertilized eggs. The GP73flox/+ mice were bred with Pdx1Cre+ mice to obtain GP73flox/+Pdx1Cre+ mice before the GP73flox/+Pdx1Cre+ mice were mated with GP73flox/+ mice to obtain the required GP73flox/floxPdx1Cre+ mice. The flox and Cre genotypes were identified by PCR. The pancreatic specificity knockout efficiency of GP73 at the protein and mRNA levels was validated by real-time quantitative PCR （qPCR） and Western blotting. The genotypes on the offspring of the crossbred mice were statistically analyzed to find out whether these genotypes conformed to the Mendel′s laws of inheritance. Additionally, the growth and development of the transgenic mice were analyzed by calculating the tissue weight or the body weight. Results GP73flox/floxPdx1Cre+ mice were obtained through mouse hybridization and identified by PCR. Compared with the control mice, the mRNA and protein levels of pancreas GP73 in GP73flox/floxPdx1Cre+ mice were significantly reduced, but the GP73 level was not significantly different in other tissues, suggesting that a proper mouse model of GP73 pancreatic-specific knockout mice was constructed. In addition, compared with the control mice, GP73flox/floxPdx1Cre+ mice had no embryonic lethality or significant difference in growth and development so that they could be used for further studies. Conclusion A GP73 pancreas-specific knockout mouse model is constructed, which can serve as a critical experimental mouse model for studying pancreatic GP73 function。
  • Original articles
    Construction of inhibitor of DNA binding 3 gene knockout human skin fibroblast cell line based on CRISPR/Cas9 system
    LUO Hong, CHEN Meihua, WANG Linxu, XU Hancui, QI Qi, PENG Hui, DUAN Xiaotao
    2023, 47(7): 493-499. https://doi.org/10.7644/j.issn.1674-9960.2023.07.003
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    Objective To construct a human skin fibroblast cell line （BJ-5ta） with stable knockout of inhibitor of DNA binding 3 （ID3） genes using the CRISPR/Cas9 technique in order to provide a cell model for elucidating the biological function of ID3. Methods In line with the biological characteristics of ID3 genes, three pairs of small guide RNA （sgRNA） were designed and inserted into LentiCRISPR v2 vector to construct recombinant plasmids. BJ-5ta cells were infected with the constructed lentiviral vector, and ID3-knockout BJ-5ta cell lines were selected through puromycin. The efficiency of ID3 gene knockout was detected by T7 endonucleaseⅠdigestion and Western blotting. The expression levels of downstream negative regulatory gene transcription products were detected via reverse transcription real-time quantitative PCR（RT-qPCR）. The effect of ID3 gene knockout on cell proliferation in BJ-5ta cells was detected using CCK-8 kit. Results T7 endonucleaseⅠdigestion and Western blotting results showed that ID3 genes in BJ-5ta cells were effectively knocked out by the CRISPR/Cas9 system. RT-qPCR results showed that the expressions of downstream negative regulatory gene transcripts were up-regulated by ID3 gene knockout. The results of CCK-8 assay showed that ID3 gene knockout had no effect on the proliferation of BJ-5ta cells. Conclusion Human BJ-5ta cell lines with ID3 knockout have been constructed, which can provide a cell model for studying the regulatory mechanism of ID3 and for gaining insights into the biological function of ID3 in human skin fibroblast cell lines.
  • Original articles
    Genomic analysis of integrating conjugative elements of Mycoplasma
    WANG Wenzhu, YUE Rui, GUO Zhiyun
    2023, 47(7): 500-507. https://doi.org/10.7644/j.issn.1674-9960.2023.07.004
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    Objective To investigate the way Mycoplasma integrating conjugative elements （MICEs）are transmitted and their impact on host Mycoplasma, and to analyze their structure, function, evolution, pangenomics, phylogenetic relationships and the molecular basis of genetic differences at the genomic level. Methods A total of 21 MICE sequences were collected, whose structural and functional characteristics were analyzed by constructing their immediate homologous gene/proteome using OrthoFinder. The genomic data of MICEs was utilized and several existing gene annotation databases were integrated to annotate the genes contained in the collected MICEs. The phylogenetic tree of the pan-genome was constructed using PanGP. The recombination events of the MICE genome were analyzed using RDP4. The positive selection sites in the MICE genome were identified using Datamonkey, the three-dimensional structure of the protein was predicted using I-TASSER, and the evolutionary relationships of MICEs were explored from a genome-wide perspective. Results Of the 419 proteins collected in the original MICEs, 386 were annotated and the key genes of their core modules were identified： T4SS type Ⅳ secretion system, SSB single chain binding protein and TraE protein. The integrated shearing module, replication module and secretion module of MICEs were also defined to make more traceable the analysis of the structure-function evolution of similar MICEs. The pan-genome of MICE exhibited a closed pattern. The topology of the core gene tree and the pangenetic tree differed significantly. Eight immediate homologous genomic recombination signals and five immediate homologous proteomic positive selection sites were identified. Results of protein structure prediction showed that these positive selection sites were associated with translocation transport of elements. Conclusion It may be inappropriate to generally describe MICEa as a class of ICEs. Our results show that the structure of MICEs not only has no core genes in the pan-genomic analysis, but also clearly exhibits several patterns in the core module and backbone analysis.
  • Original articles
    Inhibitory effect of oncolytic adenovirus rAd.DCN on the growth of non-small cell lung cancer cells
    ZHAO Huiqiang, YI Jing, DU Haitao, HAN Qiaojun, WANG Jianbin, OU Min, ZHU Yanling
    2023, 47(7): 508-514. https://doi.org/10.7644/j.issn.1674-9960.2023.07.005
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    Objective To evaluate the inhibitory effect of rAd.DCN on the growth of NCI-H292,NCI-H1703 and NCI-H1975 non-small cell lung cancer （NSCLC） cells. Methods Three types of NSCLC cells were expanded to the exponential growth stage before each type was divided into three groups： blank control group,rAd.Null treatment group and rAd.DCN treatment group. The expression of Decorin was detected by Western blotting 48 hours after infection. The survival rate of NSCLC cells infected with virus was detected by SRB method 72 hours after infection,and the killing efficiency was evaluated. At 24,48,72 and 96 hours post infection,CCK8 assay was used to measure the activity of NSCLC cells after different treatments,and the proliferation of NSCLC cells was evaluated. The expressions of target genes of Decorin were detected by quantitative real-time polymerase chain reaction（qPCR） 48 hours after infection. Results rAd.DCN could effectively infect three types of NSCLC cells and highly express Decorin protein. rAd.Null and rAd.DCN infection directly killed NSCLC tumor cells and reduced the survival rate of tumor cells,and the killing ability was related to multiplicity of infection. The killing effect of rAd.DCN was better than that of rAd.Null,and the difference in killing rates between rAd.Null and rAd.DCN on three NSCLC cells was statistically significant（P<0.05）. Compared to the control group,rAd.Null and rAd.DCN could inhibit the proliferation of NSCLC cells. Compared with the rAd.Null treatment group,the rAd.DCN treatment group had a stronger inhibitory effect on cell proliferation,and there was significant difference between the two groups（P<0.0001）. The expressions of target genes of rAd.DCN,catenin beta 1（CTNNB-1）,epidermal growth factor receptor（EGFR）,mesenchymal-epithelial transition factor（MET） and transforming growth factor-β（TGF-β） were down-regulated in NSCLC after being infected with rAd.DCN. Conclusion rAd.DCN can inhibit the growth of NSCLC cells via direct killing,inhibition of proliferation and down-regulation of target gene expressions,which is expected to become a new targeted therapy drug.
  • Original articles
    Establishment of a method for preparing cryopreserved platelets of mice and evaluating their quality
    LI Yanhong, LI Dongdong, HE Zixin, WANG Lei, SUN Zhenwei, FU Qiuxia, WANG Donggen
    2023, 47(7): 515-520. https://doi.org/10.7644/j.issn.1674-9960.2023.07.006
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    Objective To establish a preparation method for and evaluate its quality of cryopreserved platelets （CPP） of mice. Methods After the whole blood of mice was obtained by cardiac blood collection,it was anticoagulated with 14%CPDA-1 and divided into sterile centrifuge tubes. Then,platelet rich plasma （PRP） was extracted with a centrifugal force of 300×g and centrifugation time of 8 min. Furthermore,the appropriate centrifugation volume of whole blood was explored to improve the platelet recovery rate. The mouse platelets were frozen and stored at -80℃ using the method of “no washing after cryopreservation”. The quality of CPP was evaluated by platelet count,flow cytometry and platelet aggregation. Results The platelet recovery rate was the highest when the whole blood centrifuge volume was 2.0 ml,which was about 40%,and the whole blood was well stratified. Compared with fresh platelets（FP）,after CPP cryopreservation of 48 hours,there was no significant difference in platelet count （PLT）,but the mean platelet volume （MPV）,platelet distribution width （PDW）,and plateletcrit （PCT） increased. Platelet activation levels and apoptosis rate were significantly increased. The CPP aggregation rate decreased to about 40%. Conclusion A preparation method of mouse CPP is established,and the prepared mouse CPP is of good quality. The measurement indexes tend to change consistent with those of human CPP.
  • Original articles
    Effects of Chinese medicine homologous food preparations on HPO axis and HPA axis function in female rats exposed to acute cold
    JIN Lu, DONG Weiyun, BIAN Xiangyu, YANG Danfeng, LI Xi, GUO Changjiang, GAO Weina
    2023, 47(7): 521-526. https://doi.org/10.7644/j.issn.1674-9960.2023.07.007
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    Objective To observe the effects of nutritional preparations on the function of HPO axis, HPA axis and thermogenesis of brown fat in female SD rats exposed to acute cold. Methods After 3 days of adaptive feeding, 40 adult female SD rats were divided into the normal control group （C）, cold exposure group 1 （CE-1）, cold exposure group 2 （CE-2）, nutrition preparation group 1 （Nmi 1） and nutrition preparation group 2 （NMel 2）, with 8 rats in each. All the five groups were raised in a room with normal temperatures and were free to eat and drink. Group C and CE were given 2 ml distilled water every day, while group N was given 2 ml nutritional preparations daily. As the survival experiment, after 14 days of intragastric administration, the animals in the CE-1 group and NMUI 1 group were exposed to cold at -15℃ until all the experimental animals died, and the time of death was recorded. During the acute cold exposure experiment, after 14 days of intragastric administration, the animals in the CE-2 group and NMel 2 group were exposed to cold at -15℃ for 4 hours, and the anal temperature was measured immediately after the rats left the cabin. The serum, uterus, ovary and brown fat were taken, and the levels of such serum hormones as follicle stimulating hormone （FSH）, luteinizing hormone （LH）, norepinephrine （NE） and the expressions of thermogenic related proteins such as mitochondrial uncoupling protein 1 （UCP1） were detected via enzyme linked immunosorbent assay （ELISA）. The expressions of follicle stimulating hormone receptors （FSHRs）, luteinizing hormone receptors （LHRs） and uncoupling protein （UCP1） were detected by Western blotting. Results Nutritional preparations could prolong the survival time to some extent and increase the core temperature of female SD rats exposed to acute cold （P<; 0.05）. Acute cold exposure significantly up-regulated the expressions of FSHR and LHR protein in the rat ovary and decreased the serum levels of FSH and LH （P<; 0.05）, while nutritional preparation could do the opposite （P<; 0.05）. The levels of CRH, EPI and NE decreased significantly under acute cold exposure （P<; 0.05）, and nutritional preparations could change serum hormones more significantly. Acute cold exposure down-regulated the expressions of PGC-1α and UCP1 protein in brown fat （BAT） （P<; 0.05） while nutritional preparations could increase these expressions to normal. Conclusion Nutritional preparations can increase the core temperature of animals exposed to acute cold, improve the levels of HPO axis and HPA axis related hormones, regulate the expressions of HPO axis related receptors in the uterus and ovary, and promote BAT thermogenesis and cold resistance by promoting the expressions of thermogenesis related proteins.
  • Original articles
    Characteristics of formation potential of chlorination disinfection by-products of groundwater from self-contained wells and health risk assessment
    BAI Miao, JIANG Hairong, WANG Yue, XU Shaofeng, ZHANG Minglu, ZHANG Chuanfu, ZHANG Can
    2023, 47(7): 527-532. https://doi.org/10.7644/j.issn.1674-9960.2023.07.008
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    Objective To study the characteristics of formation and health risks of disinfection by-products after chlorination disinfection of self-contained wells. Methods Water samples were collected from self-contained wells in Beijing and Heilongjiang, respectively. The formation potential of disinfection by-products of the water samples was determined. Health risks were assessed according to the health risk assessment model by the US Environmental Protection Agency （USEPA）. Results For self-contained well water in Beijing, the trihalomethane formation potential （THMFP） was 15.54-72.85 μg/L, and trichloromethane accounted for 57.69% of THMFP. The haloacetic acid formation potential （HAAFP） was 0.50-6.87 μg/L, and dichloroacetic acids and trichloroacetic acids accounted for 46.56% and 37.34% of HAAFP, respectively. For self-contained well water in Heilongjiang, the THMFP was 4.34-759.58 μg/L, and trichloromethane accounted for 94.83% of THMFP. The HAAFP was 0.64-163.86 μg/L, and dichloroacetic acids and trichloroacetic acids accounted for 33.84% and 62.88% of HAAFP, respectively. The total carcinogenic risks and total non-carcinogenic risks of the formation potential of disinfection by-products from self-contained well water in Beijing were 2.02×; 10-5-9.67×; 10-5 and 3.89×; 10-2-2.27×; 10-1, respectively, which were lower than their maximum acceptable risk values by USEPA. For self-contained well water in Heilongjiang, the total carcinogenic risks of the formation potential of disinfection by-products were 5.30×; 10-6-1.05×; 10-3, and up to 40.00% （14/35） of them exceeded their maximum acceptable risk values. In addition, the total non-carcinogenic risks of the formation potential of disinfection by-products were 1.52×; 10-2-2.80, and 2.86% （1/35） exceeded the threshold limit of risk values. Conclusion For self-contained well water in Beijing and Heilongjiang, trichloromethane is the main component of THMFP, while dichloroacetic acids and trichloroacetic acids are the main components of HAAFP. Health risks mainly arise from THMFP.
  • Original articles
    Knowledge discovery of coronavirus based on MeSH terms heterogeneous networks
    CHEN Dantian, HONG Yongkai, WANG Lei
    2023, 47(7): 533-538. https://doi.org/10.7644/j.issn.1674-9960.2023.07.009
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    Objective To construct a knowledge discovery model based on MeSH terms in order to provide an effective method for discovering fine-grained knowledge associations and conduct applied research into coronaviruses. Methods MeSH terms used in literature about coronavirus were retrieved. Heterogeneous networks of MeSH terms and predicted potential associations in the networks were constructed using random walk with restart （RWR） algorithms. Finally,the prediction results were determined based on probability. Results The AUC value of ROC of the model was 0.806,and the prediction was accurate. The model predicted how to treat COVID-19 patients with acute kidney injury and how to prevent vaccination-induced thrombosis,which may be potential areas of research for treatment and prevention of coronavirus. Conclusion The model composed of MeSH terms can help discover fine-grained knowledge associations and effectively boost traditional bibliometric methods.
  • Original articles
    Design and deployment of biological technology and equipment knowledge graph system
    TONG Fan, MAO Yiqing, YANG Peixiang, LI Jiangyu, ZHAO Dongsheng
    2023, 47(7): 539-543. https://doi.org/10.7644/j.issn.1674-9960.2023.07.010
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    Objective To construct a biological technology and equipment knowledge graph in order to facilitate the research, application and teaching related to biological technology and equipment. Methods Such technologies as multi-source data collection, ontology and knowledge graph construction, text mining and analysis, knowledge annotation and curation, knowledge graph visualization and analysis were employed. Results The biological technology and equipment knowledge graph system was capable of data collection, knowledge labeling,knowledge retrival and knowledge graph presentation. Conclusion The system provides practical information and service for developing and deploying biological technology and equipment.
    • Reviews
  • Reviews
    Research progress in relationships and mechanisms between fine particles and nervous system diseases
    WAN Delian, DU Lina, SONG Lun
    2023, 47(7): 544-550. https://doi.org/10.7644/j.issn.1674-9960.2023.07.011
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    Fine particulate matter is a major component of air pollutants, which can cause significant harm to the respiratory system and cardiovascular system. In recent years, it has been found that exposure to fine particles can also affect the function of the nervous system, which is associated with the development of neurodegenerative diseases and autism spectrum disorders. Based on the data on epidemiological research, this review summarized the ways fine particles entered the central nervous system, oxidative stress, inflammatory response,changes in neuronal synaptic plasticity,and blood-brain barrier damage caused by fine particles so as to provide data for prevention of injuries to the nervous system.
  • Reviews
    Advances in macrophage recognition of pathogens and actin multimerization
    WU Feixiao, LI Shulian, YANG Guang
    2023, 47(7): 551-554. https://doi.org/10.7644/j.issn.1674-9960.2023.07.012
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    Macrophages are widely distributed in blood and various tissues, which play an important role in host immune defense. Macrophages can phagocytose and clear intracellular parasites, bacteria, tumor cells and host senescent cells. Surface receptors are essential for recognition and phagocytosis of pathogens by macrophages. Meanwhile, a variety of inflammatory cytokines are released in this process. Actin polymerization is indispensable during phagocytosis by macrophages. This paper reviews the research findings on pathogen recognition and phagocytosis mediated by receptors on macrophages.
  • Reviews
    Roles of S100A8/A9 in COVID-19 pathogenesis
    WU Shujia, JIA Yongjuan, ZHOU Kun, NI Junjun
    2023, 47(7): 555-558. https://doi.org/10.7644/j.issn.1674-9960.2023.07.013
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    Calprotectin （S100A8/A9）, an alarmin that mediates host pro-inflammatory responses during infection, regulates cytokine storm and is also involved in a variety of inflammatory diseases, such as cystic fibrosis （CF）, chronic obstructive pulmonary disease （COPD）, tuberculosis, and influenza. In recent years, some studies have found that serum S100A8/A9 levels are positively correlated with the severity of coronavirus disease 2019 （COVID-19） disease. Therefore, it is speculated that serum S100A8/A9 levels can be used as a potential biomarker of COVID-19. This paper outlines the role played by S100A8/A9 in immune response to inflammatory diseases,especially its role in the pathogenesis of COVID-19, and evaluates its potential as a diagnostic marker and therapeutic target for pulmonary infective diseases.
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