Table of Content

18 June 2021, Volume 38 Issue 3 Previous Issue   

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Research progress on the differentiation and function of human embryonic stem cells#br# DONG Yi, DONG Yigang 2021, 38 (3):  1.  doi: 10.3969/j.issn.2095-1736.2021.03.001 Abstract ( 500 )   PDF   Save The human embryonic stem cells(hESCs) are derived from preimplantation human in vitro fertilization (IVF) embryos. hESCs exhibit unique pluripotency, plasticity, unlimited capacity for self-renewal, and the ability to produce a huge number of different types of cells in vitro, so that the hESCs will open up a new way for basic research and regenerative medicine. So far, we have been able to use a variety of methods to establish and expend these cells lines, and differentiate them into specific cell lines as well. This review introduces various strategies for isolation, culture and differentiation of human embryonic stem cells into specific cell lines; then we focus on the functions of the hESC-derived neural cells in vitro study from a series of aspects: single cell, synaptic transmission and neural circuit; finally, we briefly discuss the challengesin hESCs study and its future direction. The paper aims to describe the functions of hESC from different perspectives and provide readers with the latest research progress. Related Articles | Metrics

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A summary of neurobiology salon of biological sciences SHEN Fengming, YANG Shaojie, WANG Juan, ZHU Guoqi 2021, 38 (3):  15.  doi: 10.3969/j.issn.2095-1736.2021.03.015 Abstract ( 245 )   PDF   Save The "Neurobiology Salon of Biological Sciences" was held in Hefei from October 16 to 17, 2020. The experts and representatives attending the meeting discussed the development and research status. The meeting focused on the research of neuroscience in pain, emotion, and cognition, as well as the application and transformation of traditional Chinese medicine. In these studies, optogenetics is an indispensable modern neurobiological research method. This article briefly summarized the frontier research of neurobiology in this academic salon. Related Articles | Metrics

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Amylin actions in the central nervous system and cognitive functions ZHANG Yuxiao, YAN Hongfei, YAN Chao, JI Weidong, LIU Shuai 2021, 38 (3):  20.  doi: 10.3969/j.issn.2095-1736.2021.03.020 Abstract ( 410 )   PDF   Save This article reviews amylin actions in the central nervous system in the aspects of receptor expression, signaling pathway and modulation of neuronal activity and cognitive functions. Amylin receptors are expressedin central dopaminergic system, norepinephrinergic system and serotonin system, suggesting a role in modulating the catecholamine concentrations. Amylin activates ERK, AMPK, STAT and Akt signaling pathway to change neuronal activity and modulatecognitive functions in several key brain regions. Activation of amylin signaling pathway in the central nervous system may serve as a novel therapeutic strategy for cognitive dysfunctions. Related Articles | Metrics

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Effects of ginsenoside Rg1 on depression-like behaviors, expression of hippocampal synaptic proteins and activation of glial cells in stressed mice WANG Juan, SHEN Fengming, ZHANG Zhengrong, ZHU Guoqi 2021, 38 (3):  26.  doi: 10.3969/j.issn.2095-1736.2021.03.026 Abstract ( 318 )   PDF   Save he aim of this study was to explore the effects of ginsenoside Rg1 on depression-like behaviors, hippocampal synaptic proteins and glial cell activation in stressed mice. Chronic unpredictable mild stress (CUMS) mice model was used to evaluate the effect of ginsenoside Rg1 ［10, 20, 40 mg/(kg·d), ip, 7 d］ on depression-like behaviors. The results showed that compared with control mice, sucrose preference index of CUMS mice decreased, immobility times was prolonged in the forced swimming and tail suspension experiments, which was prevented by Rg1 pretreatment. The expression of synaptic-related proteins PSD95 (postsynaptic density protein 95), Arc (activity-regulated cytoskeleton-associated protein) and BDNF (brain-derived neurotrophic factor) in the hippocampus of CUMS mice decreased, while the expression levels of astrocyte marker GFAP (glial fibrillary acidic protein), microglial marker Iba1 (ionized calcium binding adapter molecule 1) and inflammatory response factor NF-κB (nuclear factor kappa-B) increased. Administration of ginsenoside Rg1 ［20, 40 mg/(kg·d)］ increased the expression of hippocampal synaptic proteins, inhibited the activation of hippocampal astrocytes and microglia, and reduced the nuclear factor-κB (NF-κB) expression (vs CUMS， P<0.05). These data implicated that ginsenoside Rg1 prevents the depression-like behaviors likely through increasing the expression of hippocampal synaptic proteins and reducing activation of glial cells in CUMS mice. Related Articles | Metrics

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Verification of the ubiquitination of PM20D2, PABPC1 and TACO1 proteins by ubiquitin ligase E4B #br# LIU Xiangnan, FAN Yuchen, WANG Yanan, LU Yao, ZHAO Bo, WU Zhenghua 2021, 38 (3):  31.  doi: 10.3969/j.issn.2095-1736.2021.03.031 Abstract ( 228 )   PDF   Save Ubiquitin ligase E4B can transfer ubiquitin (UB) that comes from ubiquitin-activating enzyme (E1) and ubiquitin-conjugating enzyme (E2) to substrates via U-box motif. Three proteins such as peptidase M20 domain-containing protein 2 (PM20D2), polyadenylate-binding protein 1 (PABPC1), and translational activator of cytochrome C oxidase I (TACO1) ubiquitinated by E4B were verified. Total RNA was extracted from HEK293 cells and reverse-transcribed into cDNA. The cDNA was used as template for obtaining the genes of substrates by PCR, and recombinant plasmids were constructed with genes of substrates and vector, respectively. E. coli system was used to express proteins which were required for the ubiquitination of E4B substrates. Result showed that PM20D2, PABPC1 and TACO1 could be ubiquitinated by E4B.The result indicated that all the three proteins were the substrates of E4B, which would lay a foundation for the further study of the mechanism of ubiquitination in cells. Related Articles | Metrics

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Ascorbic acid promoted ex vivo proliferation of mESCs in dynamic culture TIAN Lianhua, ZHANG Tong, CAI Haibo, TAN Wensong 2021, 38 (3):  37.  doi: 10.3969/j.issn.2095-1736.2021.03.037 Abstract ( 266 )   PDF   Save In this research, the growth characteristic of mouse embryonic stem cells (mESCs) in dynamic culture was studied. With the addition of ascorbic acid, the significantly increasing time of β-galactosidase activity which was related to senescence delayed from day 6 to day 10. At the same time, intracellular reactive oxygen species level in the experimental group was lower than that in the control group, and there was a significant difference between the two groups at day 8 -day 9. Further analysis showed that the total expansion fold of mESCs and the expansion fold of SSEA-1+cells were 53.9±1.5 and 51.9±6.3 at day 8, respectively, both of which were significantly higher than those in control group without ascorbic acid, respectively (41.3±0.9 and 40.1±3.9). These data proved that ascorbic acid can reduce the level of intracellular reactive oxygen species significantly, improve the aging state of mESCs obviously and promote ex vivo expansion of mESCs. The research provides a new way to solve the aging problem of ESCs in the process of enlarged culture. Related Articles | Metrics

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Homology modeling and site-directed mutagenesis of Cassia obtusifolia trypsin inhibitor LI Chaolin, ZHANG Tian, ZOU Meng, LIAO Hai, ZHOU Jiayu 2021, 38 (3):  42.  doi: 10.3969/j.issn.2095-1736.2021.03.042 Abstract ( 267 )   PDF   Save Cassia obtusifolia trypsin inhibitor (CoTI), a member of plant Kunitz protease inhibitor family, shows strong inhibitory activity against trypsin. In this paper, the three-dimensional model of CoTI was predicted by homologous modeling. The model adopted a β-trefoil fold with β-barrel at the bottom and the lid at the top. Then, compared with the amino acid sequences and three-dimensional structures of various Kunitz protease inhibitors, a conserved Trp114 that was located in the lid region and extended to the β-barrel was found in the CoTI model. Since Trp114 formed hydrogen bonds with Leu98 and Ser137, hydrophobic interaction with Val68, Leu98 and Phe100, it was suggested to play an important role in the formation and stability of β-trefoil fold. After mutation of Trp114 residue to Ala residue using site-directed mutagenesis technique, the inhibitory activities of Trp114Ala mutant against bovine trypsin and Pierisrapae midgut trypsin were decreased by 63.4% and 62.0%, respectively. As a result, the importance of Trp114 would lay a theoretical foundation for the study of structure-function of CoTI and its related applications. Related Articles | Metrics

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Prokaryotic expression and function identification of Pichia pastoris GT1 in vitro #br# PAN Yingyue, DONG Guibin, WANG Rongbin, LIU Guoqiang, LIU Chunli, LIU Xiuxia, BAI Zhonghu, YANG Yankun 2021, 38 (3):  47.  doi: 10.3969/j.issn.2095-1736.2021.03.047 Abstract ( 202 )   PDF   Save In order to perform isothermal titration calorimetry (ITC) experimentsin vitro, pXMJ19-gt1-His recombinant plasmid was constructed.Selected by single-factor optimization experiments, the optimal host cell for the glycerol transporterGT1expression was C41(DE3); the optimal expression condition was adding 0.01mmol/L Isopropyl β-D-1-thiogalactopyranoside (IPTG) at OD600 0.7 and inducing at 30℃, 110r/min for 10h. It was identified by ultracentrifugation that some GT1 protein could be located to the plasma membrane of Escherichia coli. The detergent NP-40 with the highest resolving efficiency for GT1 was screened out. The GT1 protein was purified using nickel ion affinity chromatography with detergent buffer, and its glycerol binding affinity was measured by ITC. The results showed that the binding process was exothermic reaction, GT1 interacted with glycerol in vitro with a KDvalue of 6.58μmol/ L. The experiments showed that the GT1 protein was active and it could interact with glycerol in vitro, which provided a research basis for the study on crystal structureand function of GT1. Related Articles | Metrics

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Bioinformatic analysis and identification of Rv0357c from Mycobacterium tuberculosis TANG Wanggang, MEI Chuanzhong, GAO Jiahui, SI Yu, LIAN Chaoqun 2021, 38 (3):  52.  doi: 10.3969/j.issn.2095-1736.2021.03.052 Abstract ( 304 )   PDF   Save The aim of this work was to reveal the biological function of Rv0357c from Mycobacterium tuberculosis (Mtb) for the further exploration of a novel anti-Mtb drug target. The bioinformatic analyses of Rv0357c were conducted; the prokaryotic expression vector pET-Rv0357c for expressing Rv0357c fusion protein was subsequently constructed and introduced into Escherichia coli BL21(DE3) competent cells followed by over-production of the Rv0357c fusion protein; Rv0357c fusion protein was purified by the immobilized cobalt affinity chromatography and subjected to protein identification together with enzymatic assay. The bioinformatic analyses revealed that Rv0357c was an acidic stable hydrophilic protein containing two conserved motifs and conserved active site residues observed in previously identified adenylosuccinate synthetases (AdSSs), and it showed low sequence identities(＜40%) to human enzymes (muscle isoform, AdSS1, and non-muscle isoform, AdSS2). Based on the SDS-PAGE analysis, the Rv0357c fusion protein was found to be expressed as inclusion bodies with an apparent subunit molecular mass of 48 ku, and the target enzyme in soluble form could be obtained after solubilizing the inclusion bodies, affinity chromatography and subsequent protein refolding with a refolding yield of approximate 63%. The overproduced protein obtained here was further identified as the target protein as demonstrated by Western Blot analysis using Anti-6×His antibody. Furthermore, enzymatic assay revealed that the biological activity of AdSS could be detected using refolded Rv0357c fusion protein with a specific activity of 0.016 1 U/mg. In summary, the Rv0357c is a functional enzyme with AdSS activity. This work will lay an experimental foundation for the further functional identification and the development of a new drug target against Mtb. Related Articles | Metrics

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Research on SNRPD1 as a novel breast cancer prognostic marker #br# CHEN Xiao, DAI Xiaofeng 2021, 38 (3):  57.  doi: 10.3969/j.issn.2095-1736.2021.03.057 Abstract ( 320 )   PDF   Save SNRPD1is responsible for encodinga protein belonging to the spliceosome family, which is recognized as an essential player in RNA splicing process. It was later found to be implicated in the development of several types of cancer. We used bioinformatics and molecular assays to predict and validate the function ofSNRPD1in breast cancer development. Transcriptome data analysis revealed that higherSNRPD1expression was observed in tumor tissue compared to normal tissue andSNRPD1was highly expressed in Triple Negative subtype, the most malignant subtype of breast cancer. Survival analysis confirmed that patients with higherSNRPD1expression exhibits worse survival rate and higher relapse probability. Correlation analysis showed thatSNRPD1co-expressing genes are majorly involved in cell proliferation pathway and cell division process. Molecular assays confirmed silencingSNRPD1will lead to reduced cell proliferation and under-regulated JNK expression. Our study revealed the function ofSNRPD1in breast cancer development and proposesSNRPD1as a potential breast cancer prognostic marker, which can deliver valuable insights for breast cancer clinical treatment. Related Articles | Metrics

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Knockout and identification of brain type aromatase gene cyp19b in medaka CHANG Yuyang, WU Xiaowen, GUO Haiyan, GUAN Guijun 2021, 38 (3):  63.  doi: 10.3969/j.issn.2095-1736.2021.03.063 Abstract ( 266 )   PDF   Save Aromatase is the key enzyme for the synthesis and conversion of estrogens from androgens.Brain type aromatase(cyp19b) is predominantly expressed and displays a sexual dimorphism in adult brain, in contrast to a slight expression with no sexual dimorphism in adult gonad of medaka (Oryzias latipes). In order to explore the bio-function ofcyp19bgene in brain and gonad development, we used CRISPR/Cas9 system to targeted knockoutcyp19b. Two targeting sites were designed at exon 2 and exon 4, respectively.Microinjection of aliquots containing gRNAs and Cas9 protein was carried out into 1-cell fertilized eggs. Results showed that two heritablecyp19bmutant alleles were successfully obtained, and twocyp19bknockout mutant homozygous lines were selected through heterozygous mating, genomic PCR and DNA sequencing analysis. There was no significant male-or female-bias in both lines ofcyp19bknockout homozygotes. In both lines ofcyp19bknockout homozygotes, the embryo could develop normally and the adult could reproduce normally, suggesting that the loss ofcyp19bgene might not directly affect the sexual differentiation of gonads. Related Articles | Metrics

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Phylogenetic, molecular evolution and expression analyses of HMGR gene in loblolly pine MAO Jipeng, DU Chengju, HUANG Linwang, LIU Tianyi, HUANG Shaowei 2021, 38 (3):  68.  doi: 10.3969/j.issn.2095-1736.2021.03.068 Abstract ( 266 )   PDF   Save The 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) is a key enzyme in the MVA biosynthesis pathway of terpenoids. To understand the phylogenetic of its coding gene HMGR and its molecular evolution and expression characteristics in the clade of loblolly pine, in this study, 19 representative species of HMGR sequences were selected for phylogenetic and molecular evolution analysis, and 5 different tissue parts of loblolly pine were used as materials to analyze the expression characteristics of HMGR. The results showed that the HMGR was relatively conserved in the evolution process. The average genetic distance between sequences was 0.323, and the average selection pressure was 0.1212, which indicated that HMGR was mainly subjected to the purifying selection during the evolution process. The selective pressure was 0.4293 in the clade of loblolly pine. Fourteen sites were detected to be positive selective, and the value of posterior probability was greater than 0.95, which indicated that the different selective pressure on each site of HMGR was significant. HMGR had the highest expression level in young leaves of loblolly pine, showing the tissue specific expression of HMGR. Related Articles | Metrics

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Effects of two elicitors on disease resistance and rhizosphere bacterial community of tomato #br# SU Yingying, JIN Zhaoxia, CUI Yuxing 2021, 38 (3):  73.  doi: 10.3969/j.issn.2095-1736.2021.03.073 Abstract ( 156 )   PDF   Save Tomato grey mould is a worldwide fungal disease caused by Botrytis cinerea. Induced resistance has the advantages of environmental protection and sustainability. In this paper, Pseudomonas choloeaphtis and β-amino butyric acid with potential for resistance were used to determine the anti-disease effect of tomato gray mold and the regulation of rhizosphere bacterial community structure by two pretreatments. The pot test and real-time fluorescence quantitative technology were used to obtain the resistance to gray mold and tomato resistance gene expression under different pretreatments. The 16S rDNA high-throughput sequencing was used to investigate the rhizosphere bacterial community structural change. The results showed that both Pseudomonas choloeaphtis and β-aminobutyric acid treatment could reduce the incidence of tomato plants with reducing the disease index by 60.4% and 36.6% respectively, promoted the expression of resistance genesNPR1andPR1, and stimulated the SA signaling pathway. High-throughput sequencing results showed that pretreatment increased the abundance of Proteobacteria, Pachyphyta and Bacteroides in rhizosphere bacteria, adjusted the abundance and structure of the dominant genus of tomato rhizobacteria. The abundance of probiotic flora in rhizosphere increased significantly. Related Articles | Metrics

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Effects of different gut microbes on mating behavior of Drosophila melanogaster WEI Bofan, WANG Lu, LI Xiaozhe, LI Xiaohui, LI Miaomiao, QIAO Huili 2021, 38 (3):  78.  doi: 10.3969/j.issn.2095-1736.2021.03.078 Abstract ( 278 )   PDF   Save The germ free and gnotobioticDrosophilamelanogastersystem was constructed, and the effects of different gut microbes on the mating behavior of germ free and conventionalD. melanogasterwere studied. The fertilized eggs ofD. melanogasterwere sterilized in vitro and cultured in sterile condition to obtain germ freeDrosophila.Lactobacillus plantarum,Acetobacterium malorumandSaccharomyces cerevisiaewere inoculated in fly culture medium, respectively, then the germ free and conventional flieswas kept respectively in the culture medium for a certain period of time to obtain different gnotobioticDrosophila.All flies were dissected and the total RNA of the gut was extracted and reverse transcribed, and the abundance of different microbes in the gut of germ free and conventional flies was detected by quantitative real-time PCR (qRT-PCR). Meanwhile, single pair mating assays were carried out to compare the copulation success rate and copulation latency of different treated flies. The results showed that the germ free and gnotobioticDrosophilasystem was successfully constructed, and the specific microbe was successfully enriched in the gut of the germ free and conventional flies after co-cultivation with the single microbe, andL. plantarumcould affect the mating behavior ofDrosophilaby prolonging the copulation latency. Related Articles | Metrics

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Effects of drought stress on photosynthesis of upland-island RIL population at blossom and boll stage #br# IANG Menghui, SUN Fenglei, FAN Rong, SHI Yingying, LIU Yali, ZHAO Keke, QU Yanying, CHEN Quanjia 2021, 38 (3):  83.  doi: 10.3969/j.issn.2095-1736.2021.03.083 Abstract ( 197 )   Save The F2∶6 upland-island RIL population of upland cotton (Zhong 07) and island cotton (Xinhai 20) was used as the materials, and was treated by a field drought-stress method at the blossom and boll stage. After the measurement of photosynthetic traits, the drought resistance of upland-island RIL population was evaluated through the methods of variance analysis, principal component analysis, cluster analysis and integrated drought assessment system. The results showed that the response degree of each indicator was different in different cotton materials under drought stress at blossom and boll stage, among which, the A, E and WUE showed significant changes. Through D value clustering analysis, the cotton materials were divided into four groups, 21 materials in group Ⅰ with strong drought resistance type, 7 materials in group Ⅱ with drought resistance type, 26 materials in group Ⅲ with drought sensitive type and 16 materials in group Ⅳ with extreme drought sensitive type. Through cluster analysis, the cotton materials of more drought-resistant lines than that of Zhong 07 and more drought-sensitive lines than that of Xinhai 20 were obtained. Related Articles | Metrics

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Variation in three dominant plant species leaf traits and comprehensive evaluation under heavy metal pollution in mining areas CAO Jingxiao, LU Sufen, WEI Yansong, WEI Lianqiang, HUANG Qitang 2021, 38 (3):  88.  doi: 10.3969/j.issn.2095-1736.2021.03.088 Abstract ( 182 )   PDF   Save In this study， three dominant species from mining area of Nandan county including Thysanolaena latifolia(Roxb.) Honda, Neyraudia reynaudiana (kunth.) Keng and Miscanthus sinensis Anderss were selected to investigate phenotypic traits in respond to the heavy metal pollution (lead and cadmium) by means of variance, variation coefficient and correlation analysis. The results showed that variation in population (80.65%, 64.25% and 76.17%) was greater than that among populations (6.79%, 24.30% and 15.57%) among three dominant plant species, the average differentiation coefficients of phenotypic traits were 91.64%, 72.24% and 83.83%, respectively, and the variation in population was the variation of three dominant plant species.The average variation coefficients of the three dominant plant species were 16.10%, 19.31% and 22.13%, respectively, and their variation ranges were 8.86%-25.43%, 6.46%-34.82% and 11.49%-36.39%, respectively. The M. sinensis showed the lowest SLA(110.75±23.69) cm2/g and the middle level LDMC(0.37±0.06) g/g; the N. reynaudiana showed the highest SLA(139.89±10.66) cm2/g, the middle level LDMC(0.39±0.03) g/g, and the T. maxima showed the highest SLA(142.98±40.41) cm2/g and the middle level LDMC(0.37±0.04) g/g. The pH value, water content and TP of soil were significantly correlated with significant characters of T. maxima and M. sinensis, while phenotype traits of N. reynaudiana was not significantly correlated with eight soil factors. Related Articles | Metrics

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Research progress of biotin metabolism regulators in Prokaryotes CHEN Yuhong, WU Hang, ZHANG Buchang 2021, 38 (3):  94.  doi: 10.3969/j.issn.2095-1736.2021.03.094 Abstract ( 405 )   PDF   Save Biotin (also called vitamin H or B7) is an important micronutrient that can participate in various metabolic processes as a cofactor for biotin-dependent enzymes (biotin-dependent carboxylase, decarboxylase, and transcarboxylase). When biotin is deficient, the vitality of the cells will be reduced, and even death will occur. The earliest discovered biotin metabolism regulator is BirA, which contains not only the DNA binding domain, but also the functional domain of protein biotin ligase.In recent years, it has been found that the biotin metabolism process in Prokaryotes presents a variety of regulatory modes. Currently, three types of regulatory factors, suchas BirA, BioR, and BioQ, have been identified, which respectively play a transcriptional regulatory role in the biotin metabolism genes of Prokaryotes such as γ-Proteobacterias, α-Proteobacterias and Actinomycetes. This review summarized these three types of regulatory factors, and focused on the research progress of their regulatory functions so as to provide a theoretical guidance for further revealing the molecular mechanisms of these regulatory factors. Related Articles | Metrics

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Research in lignin peroxidase LIANG Xiaoyu, CUI Zhoulei, WANG Hongcheng, CHEN Huayou 2021, 38 (3):  99.  doi: 10.3969/j.issn.2095-1736.2021.03.099 Abstract ( 611 )   PDF   Save This paper reviewed the related research progress of the catalytic mechanism, enzyme activity assay, cloning and expression, synergy, application of lignin peroxidase. Lignin peroxidase has broad application prospects in chemical industries, ecological restoration and biological feed industries, however, the industrialized application of lignin peroxidase is limited by its production and enzyme activity. The key to promote the industrialized application of lignin peroxidase is to increase the enzyme expression, improve the enzymatic properties, and explore the efficient use of lignin peroxidase, which is also a research hotspot in recent years. The results provide references for further research and development of lignin peroxidase， and a forecast of the research prospect of lignin peroxidase. Related Articles | Metrics

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Advances in RASSF4 gene promoter methylation and tumors #br# YAN Yanan, CHEN Xingzhen, ZHOU Cefan, TANG Jingfeng 2021, 38 (3):  103.  doi: 10.3969/j.issn.2095-1736.2021.03.103 Abstract ( 236 )   PDF   Save Ras association domain family member 4 (RASSF4) involves in multiple cell physiological activities such as cell proliferation, differentiation and apoptosis. The disordered expression of RASSF4 had shown significant correlation with tumors. To address this issue, the molecular structure, biological function, the relationship between promoter methylation and tumors were reviewed. One of the molecular mechanisms regulatingRASSF4gene expression is the DNA methylation of its gene promoter. However, the association and significance of the DNA methylation ofRASSF4gene promoter with the development of tumors are not clear. The application prospect of demethylation treatment of tumors was discussed. This review aimed to clarify the biological profile and role of RASSF4 in various tumors, thus providing a theoretical basis for the development of RASSF4 as a detection and treatment target for specific tumors. Related Articles | Metrics

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Prokaryotic expression of soluble B lymphocyte stimulating factor and preparation of anti-BLys monoclonal antibody aginst sBLyS LIU Chong, GAO Zhaowei, WANG Huiping, HE Ting, LIU Li, DONG Ke 2021, 38 (3):  108.  doi: 10.3969/j.issn.2095-1736.2021.03.108 Abstract ( 192 )   PDF   Save To develop a soluble B lymphocyte stimulating factor (sBLyS) detection assay, the sBLyS encoding gene was cloned in this study, and sBLyS recombinant protein was expressed by using Escherichia coli BL21(DE3). The recombinant protein was purified by nickel column affinity chromatography. BALB/c mice were immunized with purified sBLyS recombinant protein, and cell lines secreting anti-BLyS monoclonal antibody (mAb) were obtained by using cell fusion technology. The mAb was purified by using n-Caprylic acid combined ammonium sulfate precipitation assay, and Western Blot and ELISA were used to analyze the activity and titer of the mAb. The unlabelled mAb and the HRP labeled mAb was used as the capture antibody, and the detection antibody, respectively, and then the double antibody sandwich ELISA test were performed to screen the optimal combination. The results showed that the sBLyS recombinant protein was expressed and purified. Four cell lines secreting the sBLyS monoclonal antibody hybridoma were obtained (named 2E4, 2H8, 4C5 and 4C12) with the titer of 9.6， 10.8, 20.4 and 26.6ng/mL, respectively. Western Blot results showed that the purified monoclonal antibodies had sBLyS protein binding activity. Double antibody sandwich ELISA showed that the combination of 2H8 and HRP-4C5 could be used to detect sBLyS, and the testing linearity ranged from 0.4ng/mL to 1.6ng/mL (R2=0.995). Related Articles | Metrics

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Construction and reform of physiological experiment course for biotechnology major #br# LENG Ye 2021, 38 (3):  112.  doi: 10.3969/j.issn.2095-1736.2021.03.112 Abstract ( 234 )   PDF   Save In physiology teaching, experiment course plays an important role. Based on the original "physiological experiment" teaching, combined with the professional training direction of stem cell and regenerative medicine, basis and translational medicine research of major disease, this paper carried out teaching reform. The contents of traditional physiology experiment were redesigned, new experimental items were introduced, new experimental teaching materials and operation videos were compiled, the traditional experimental teaching methods were reformed, the design and implementation of innovative and exploratory experiments were encouraged, and the ideological and political contents were integrated into the experimental courses to enhance the experimental effect. Through the teaching reform, the experimental courses were more in line with the talent training objectives of the college, and more closely combined with the theoretical courses. The experimental contents were more novel, scientific and practical. It focused on the cultivation of students′ practical ability and independent thinking ability, which was conducive to the students′ understanding of the theoretical course and the improvement of their scientific research ability. Related Articles | Metrics

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An exploration on curriculum politics practice in the courses of life sciences WU Genfu, WU Kejie 2021, 38 (3):  116.  doi: 10.3969/j.issn.2095-1736.2021.03.116 Abstract ( 604 )   PDF   Save Curriculum politics is the practical activities of ideological education relying on professional or general courses. The curriculum politics in the courses of life sciences should be carried out based on discipline characteristic by taking general courses as a carrier, and social hot spots, major scientific achievements and discipline history as the starting points. In view of the current limited class hours and insufficient political abilities for professional courses, it is suggested that a general course named "Life Education" is established by integrating all of the cases suitable for ideological education in life and healthy courses. The contents such as life and life education, the origin and evolution of life, the diversity and uniqueness of life, the sacredness and rarity of life, the fragility and tenacity of life, the harmony and responsibility of life, and the value and significance of life are suitable to be the key points for the curriculum. By utilizing the outstanding scientific achievements, especially the Nobel Prize achievements, the goal of imparting knowledge and educating people is achieved. Related Articles | Metrics

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Exploration and practice of "internet + teaching" in plant biology course CAI Chong, ZHU Cheng 2021, 38 (3):  120.  doi: 10.3969/j.issn.2095-1736.2021.03.120 Abstract ( 233 )   PDF   Save From the perspective of characteristics of the subject and taking the national first-class undergraduate course as an example, this paper summed up some specific problems and measures about teaching content arrangement and teaching methods improvement combined with years of teaching practice of the authors themselves to provide some teaching references, so as to improve the teaching quality and learning effect of the course．"Internet +" has given birth to new educational productivity. Through the deep integration of information technology and classroom teaching and effective interaction between online and offline, the course of plant biology realized the diversification of classroom experience, guided the change of students' learning style, formed diversified academic evaluation and promoted the continuous improvement of teaching activities. The course construction of plant biology made fruitful exploration in adapting to the learning direction of contemporary students. It is expected to jointly promote the construction and application of first-class courses and integrate into the innovation and development of teaching and learning in the new era. Related Articles | Metrics

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Teaching reform and practice of entomology experiment for plant protection specialty GAO Ping, DONG Hui, XUE Caiyun, LU Ying, WANG Hongping 2021, 38 (3):  123.  doi: 10.3969/j.issn.2095-1736.2021.03.123 Abstract ( 220 )   PDF   Save n order to adapt with the needs of cultivating innovative talents and improve the shortcomings of traditional entomology experiment course in the implementation process, teaching reform of entomology experiment for plant protection specialty was carried out. A multi-level experimental teaching system of entomology was constructed by adjusting and optimizing teaching content; student-centered experimental teaching mode was established by enriching teaching methods and improve teaching modes; the experimental teaching conditions were improved by strengthening the construction of insect specimens and hardware and optimizing laboratory management; the evaluation system of experimental teaching of entomology was also improved through establishing diversified assessment methods. The reforms have achieved good results in improving the quality of experimental teaching, mobilizing students' learning enthusiasm and improving their practical ability and innovation ability. Related Articles | Metrics

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Design and practice of formative assessment of PBL-SPOC integrated teaching model for food microbiology#br# TAN Haigang, LI Jing 2021, 38 (3):  127.  doi: 10.3969/j.issn.2095-1736.2021.03.127 Abstract ( 215 )   PDF   Save The reform of teaching assessment methods is crucial for the teaching effects of course and the quality of talent cultivation. In order to improve the innovative ability, the independent learning and thinking abilities of students, the small private online course (SPOC) and problem-based learning (PBL) teaching mode were introduced in the teaching of food microbiology. The quality of teaching assessment and feedback had a significant effect on the practice of the PBL-SPOC integrated teaching model. Based on the shortcomings of summative assessment under the background of MOOC, the advantages of formative assessment in promoting teaching quality, the characteristics of online and offline hybrid teaching and food microbiology, the three-stage formative assessment in the pre-class preview, the class teaching and the after-class review of PBL-SPOC integrated teaching model of food microbiology were designed, practiced, discussed and summarized. It provided references for the practice of formative assessment in professional basic courses, the goal achievement of teaching feedback, the independent learning ability cultivation and the reform of assessment methods. Related Articles | Metrics