18 October 2023, Volume 40 Issue 5 Previous Issue   
The killing mechanism of lung cancer cells by nonthermal atmospheric pressure plasma mediated by ROS
ZHOU Yuanyuan, LIU Fang, CHEN Jinwu, LIU Lei, SONG Wencheng, CHU Yannan
2023, 40 (5):  1.  doi: 10.3969/j.issn.2095-1736.2023.05.001
Abstract ( 239 )   PDF  
The nonthermal atmospheric plasma (NAP) was used to treat lung cancer cells H460. With the extension of NAP treatment time, MTT assay showed that NAP could decrease the cell viability of H460 significantly, JC-1 assay showed that NAP decreased the mitochondrial membrane potential of H460, which represented the occurrence of cell apoptosis. Western Blot experiment showed that NAP inhibited the expression of PI3K, AKT, JNK and STAT3, demonstrating the killing effect of NAP related to the expression of PI3K/AKT and JNK/STAT3/AKT. Besides, the addition of ROS scavenger NAC effectively eliminated the damage caused by NAP, suggesting that the killing of lung cancer H460 cells by NAP were mediated by ROS, which provided theoretical basis for the feasibility of NAP in the treatment of lung cancer.
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Glutamine synthetase regulates autophagy to improve radiotherapy sensitivity in hepatocellular carcinoma cells
HE Yuan, QIAN Junchao
2023, 40 (5):  6.  doi: 10.3969/j.issn.2095-1736.2023.05.006
Abstract ( 93 )   PDF  
The aim of this study was to investigate the relationship among radiotherapy sensitivity, glutamine synthetase (GS) and autophagy in human hepatocellular carcinoma cells. After X-ray irradiation, CCK-8 and colony formation assay were used to measure the proliferation of HepG2 cells treated with GS inhibitor (L-methionine sulfonimide, MSO). Real-time fluorescence quantitative PCR and protein immunoblotting assays (Western Blot, WB) were used to detect the effect of MSO on GS expression, and GS activity assay kits were used to detect the efficiency of inhibition of GS activity by MSO. Monodansulfonyl cadaverine (MDC) staining as well as WB assay techniques were used to detect intracellular autophagy to determine the effect of IR and MSO on autophagy. The results indicated that MSO caused an increase in intracellular GS expression, but GS activity was significantly inhibited. In addition, MSO reduced the proliferative capacity of cells after radiotherapy and inhibited radiotherapy-induced autophagy. Results suggested that MSO could enhance the sensitivity of HepG2 cells to radiotherapy by inhibiting GS activity, which was closely related to its regulation of autophagic flow.
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HIGD1A regulates autophagy to enhance the radiation resistance of HeLa cells
ZHAO Xipeng, ZHAO Guoping
2023, 40 (5):  11.  doi: 10.3969/j.issn.2095-1736.2023.05.011
Abstract ( 74 )   PDF  
This article mainly explored the mechanism of mitochondrial protein HIGD1A enhancing the radiosensitivity of cervical cancer cells (HeLa) by regulating autophagy. Using control cells and HIGD1A knockdown HeLa cells as research subjects, cells were treated with autophagy inducer Earle’s balanced salt solution (EBSS) and ionizing radiation. The expression levels of cell autophagy marker proteins LC3 and P62 were detected by Western Blot, and the fluorescence expression level of Ad-mCherry-GFP-LC3B recombinant adenovirus infected cells was detected. Cells were irradiated after the pretreatment of chloroquine/rapamycin for 2 hours, then their viabilities were detected using CCK-8 48 hours later. The expression levels of caspase-7 and cleaved caspase-3 proteins were detected by Western Blot after rapamycin treatment. The results showed that compared with the control cells, knocking down HIGD1A inhibited the generation of LC3, P62 degradation, and the generation of yellow and red spots in mCherry-GFP-LC3B after EBSS or ionizing radiation treatment. CCK-8 assay results showed that knockdown of HIGD1A combined with chloroquine treatment reduced cell viability after ionizing radiation treatment. In addition, knocking down HIGD1A significantly increased the expression levels of caspase-7 and cleaved caspase-3 proteins in tumor cells, and there were still high levels of apoptosis in HIGD1A knocked down cells after rapamycin treatment. The results indicated that HIGD1A was involved in regulating the radiation sensitivity of tumor cells through the autophagy pathway.
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Gene synthesis, prokaryotic expression of human interferon ε and its characterization by mass spectrometry
YANG Yinhe, DI Bin, YANG Dasong
2023, 40 (5):  16.  doi: 10.3969/j.issn.2095-1736.2023.05.016
Abstract ( 65 )   PDF  
First of all, the gene of human Ifn-ε was synthesized, the expression vectors of Ifn-ε/pET-21a(+) and Ifn-ε/pET-22b(+) were constructed respectively after codon optimization. The expression vector was transformed into Escherichia coli and the expression conditions were further optimized. Two dominant strains Ifn-ε/pET-21a/BL21A and Ifn-ε/pET-22b/BL21A were obtained. At last, IFN-ε with high purity was isolated from Ifn-ε/pET-22b/BL21A throgh fermentation, extraction, purification, and its structure was confirmed by mass spectrometry. An effective expression system of human IFN-ε has been established to provide large quantity of protein for the study of its structure and mechanism. Moreover, the establishment of expression system through gene synthesis laid the foundation for the study of structure-activity relationship by amino acid mutation.
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Cathepsin D contributes to SRBSDV replication in white-backed planthopper
FU Yating, GUO Dongyang, WU Qingfa
2023, 40 (5):  20.  doi: 10.3969/j.issn.2095-1736.2023.05.020
Abstract ( 76 )   PDF  
When the white-backed planthopper (Sogatella furcifera) acquires Southern rice black streaked dwarf virus(SRBSDV) by feeding on SRBSDV-infected rice, the SRBSDV binds to receptors on the cell membrane and enters the host cell via clathrin-dependent endocytosis. Cathepsin D (CathD) is an aspartic protease, which is involved in the process of endosome/lysosomal protein degradation and viral release into the cytoplasm. However, it is unclear whether CathD plays a role in SRBSDV infection of white backed planthopper. RNA interference (RNAi) was used to silence the expression of CathD gene in the white-backed planthopper. The results showed that knocking down the mRNA expression level of CathD in the white-backed planthopper and subsequently SRBSDV feeding did not affect the survival rate of the white-backed planthopper, however, the viral carrier rate and viral quantity in the white-backed planthoppers were decreased. Meanwhile, knockdown the expression of the homologous CathD gene in Drosophila cells decreased DCV replication significantly compared with that of the control group. The results demonstrated that CathD may assist virus proliferation and replication in insects using a conserved mechanism, suggesting that CathD might be a potential target protein for the prevention and control of SRBSDV disease.
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Study on the infection of Caenorhabditis elegans by Colletotrichum gloeosporioides
XIAO Yue, YI Xingying, ZHANG Donghua, LIU Li, YAN Xiaohui, WU Jianrong
2023, 40 (5):  24.  doi: 10.3969/j.issn.2095-1736.2023.05.024
Abstract ( 55 )   PDF  
The effects of the plant pathogen on the basic biological characteristics of Caenorhabditis elegans were studied by co-culture of C. elegans and C. gloeosporioides. The results showed that C. gloeosporioides had great impact on the survival, body length and reproductive of C. elegans. The survival rate of C. elegans decreased obviously after exposing to C. gloeosporioides, and the age of the worm was directly related to the half death time, which was 108 h in L1 stage and 144 h in L4 stage, respectively. The conidia concentration of C. gloeosporioides spores was negatively correlated with the survival of C. elegans. The body length of C. elegans in co-culture with C. gloeosporioides grew shorter evidently, and the reproductive of C. elegans was significantly decreased. However, C. gloeosporioides had no significant influence on C. elegans’ motility capacity and pharynx pump capacity. The results indicated that C. gloeosporioides could harm C. elegans to some extent and might activate the innate immune system of C. elegans. The interaction system between C. gloeosporioides and C. elegans may be a powerful tool for studying the evolution of innate immunity.
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Isolation and screen of endophytic fungi from Peucedanum praeruptorum and secondary metabolites from strain bhqh-2
HANG Shikai, XU Qing, LIU Lin, YANG Zhengyou, ZHAO Fengchun
2023, 40 (5):  29.  doi: 10.3969/j.issn.2095-1736.2023.05.029
Abstract ( 51 )   PDF  
This study aims to isolate endophytes with good bioactivity from Peucedanum praeruptorum to provide a basis for the research and development of new drugs. The endophytic fungi were isolated by tissue block method and fermented in ME medium. The fermented liquids were extracted by ethyl acetate and concentrated to obtain the crude extracts. The antimicrobial activities of the crude extracts against four bacteria, Staphylococcus aureus, Salmonella enteritidis, Pseudomonas aeruginosa and Escherichia coli, and four fungi, Fusarium graminearum, Verticillium dahliae, Botrytis cinerea and Candida albicans, were determined by cup-plate method. The cytotoxicity against human lung cancer cell line A549 and human breast cancer cell line MDA-MB-231 was determined by MTT method. Finally, morphological observation combined with ribosomal internal transcription spacer sequence (rDNA-ITS) was used to identify one strain with potent activity, and the secondary metabolites of the strain were isolated, purified and identified. A total of 7 strains of endophytic fungi were isolated. Among them, 4 strains, bhqh-2, 3, 5 and 7 showed good biological activities, and their crude extracts had inhibitory effects on all of the test bacteria and fungi, and the inhibitory rates on the two kinds of cancer cells were more than 95%. Strain bhqh-2 was identified as Aspergillus aspergillus, and 6 known compounds were isolated and purified from its crude extract. In conclusion, some P. praeruptorum associated fungi showed good biological activity, which provided a basis for the study of endophytic metabolites and their biological activities.
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The relationship between arbuscular mycorrhizal fungi (AMF) and anthracnose occurrence in Camellia oleifera
YANG Yalin, WU Ziqiang, LIU Li, ZHANG Donghua, MA Huancheng, WU Jianrong
2023, 40 (5):  35.  doi: 10.3969/j.issn.2095-1736.2023.05.035
Abstract ( 54 )   PDF  
In order to control Camellia oleifera anthracnose with arbuscular mycorrhizal fungi (AMF), the effects of AMF on Camellia oleifera anthracnose were investigated. The parameters such as AMF spore density, richness, diversity index, mycorrhizal colonization rate of C.oleifera and the disease occurrence were studied by wet screening precipitation and AMF colonization staining methods. Results showed that there were significant differences in AMF spore density and colonization rate between diseased and healthy C.oleifera roots. The spore density, richness and colonization rate of AMF were negatively correlated with the incidence rate and disease index of C.oleifera anthracnose, and the diversity index was also negatively correlated with the incidence rate and disease index of C.oleifera anthracnose. It showed that the higher the spore density, mycorrhizal colonization rate, richness and diversity index of AMF, the less the anthracnose of C.oleifera.
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Effects of adding photosynthetic bacteria to microalgae diets on the development and reproduction of Pseudodiaptomus annandalei #br#
PAN Yao, LI Feng, GAO Menghan, PENG Ke, RUI Xiangyu, LI Changling, HUANG Xianghu
2023, 40 (5):  41.  doi: 10.3969/j.issn.2095-1736.2023.05.041
Abstract ( 42 )   PDF  
In order to explore the effects of photosynthetic bacteria on the growth and reproduction of copepods, this paper studied the effects of different microalgae diets on the development and reproduction of Pseudodiaptomus annandalei. The results showed that different types of bait microalgae had significant effects on the development and reproduction of Pseudodiaptomus annandalei (P<0.05). When fed with Isochrysis zhanjiangensis, Pseudodiaptomus annandalei developed fast with the highest survival rate and the highest total number of larvae. Subsequently, different concentrations of photosynthetic bacteria were added to the Isochrysis zhanjiangensis. The results showed that when the concentration of photosynthetic bacteria reached 1 mgC/L with the same amount of microalgae, the development time of nauplii and copepods of Pseudodiaptomus annandalei was shortened by 20% and 14.5%, respectively, the survival rate of individuals was increased by 21.7% and the total number of larvae increased by 20.4%. However, when the concentration of added bacteria was 2 mgC/L, the growth and the reproduction of Pseudodiaptomus annandalei were significantly inhibited. The results showed that adding an appropriate amount of photosynthetic bacteria to the microalgae feed was beneficial to the development and reproduction of Pseudodiaptomus annandalei.
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Characteristics and influencing factors of metazooplankton functional groups in Qingcaosha Reservoir
HONG Chencong, HU Quan, CHEN Haoruo, XU Saisai, ZHANG Jian, CHEN Lijing
2023, 40 (5):  47.  doi: 10.3969/j.issn.2095-1736.2023.05.047
Abstract ( 60 )   PDF  
To analyze the characteristics of metazooplankton functional groups and their relationship with water environmental factors in Qingcaosha Reservoir, a monthly sampling survey was conducted in 2021. A total of 47 metazooplankton species were collected and divided into 9 functional groups. The average annual biological density of metazooplankton was 82.4 ind/L, and the average annual biomass was 0.3460 mg/L. Rotifers carnivore(RC), middle copepods and claocera filter feeders(MCF) and large copepods and claocera filter feeders(LCF) constituted the main part of the total zooplankton biomass in Qingcaosha Reservoir. Pearson analysis showed that small copepods and claocera carnivore(SCC) had a significant correlation with middle copepods and claocera carnivore(MCC) and small copepods and claocera filter feeders(SCF) (P<0.05), and the functional group SCF, SCC, MCF, and MCC had a significant correlation (P<0.01), while RC and rtifers filter feeders(RF) showed no significant correlation (P>0.05). Redundancy analysis showed that in addition to functional group RC, LCF and LCC, other functional groups were significantly negatively correlated with water depth and dissolved oxygen, and positively correlated with water temperature. Total dissolved solids and ammonia nitrogen were significantly positively correlated with functional group RC and LCC, and significantly negatively correlated with functional group LCF.
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Comparative analysis on otolith morphology of Selaroides leptolepis and Decapterus macrosoma in the southcentral South China Sea
LI Weichang, ZHU Guoping, WANG Xuehui, LIN Longshan, LI Yuan, DU Feiyan
2023, 40 (5):  54.  doi: 10.3969/j.issn.2095-1736.2023.05.054
Abstract ( 38 )   PDF  
In order to understand the morphological characteristics of sagittal otoliths of fish species in Carangidae and to study the effectiveness of different machine learning algorithms on the population classification of Carangidae in the South China Sea, otolith samples of Selaroides leptolepis and Decapterus macrosoma were collected from the South China Sea. The otoliths were measured with 4 basic morphological parameters and transformed into 6 morphological indexes, and the differences in otolith morphology were compared between two species, forty four elliptic Fourier descriptor coefficients were extracted from the otoliths for principal component analysis (PCA), and four different machine learning classification models, namely, linear discriminant analysis (LDA), random forest(RF), K-nearest neighbor(KNN), and support vector machines(SVM) were used to discriminate them. The result showed that the otolith parameters and body length of the two species were significantly different. The length, height, area, perimeter, and body length of the two species were all power functions. From the analysis of morphological indexes, the otoliths of S. leptolepis had a lower ring rate described as more round and more regular, there were significant differences in morphological indexes between the two groups. PCA showed that the first and second principal component accounted for 20.1% and 13.3% of the total variation, respectively. Among the four classification models, the highest correct rate of RF was 100%, and the lowest was 93.3% for SVM.
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Niche change and interspecific association of dominant phytoplankton species in Wuxikou
HUANG Xueping, CHEN Ziyue, HE Guoqing, LIAO Weidong, YAO Yuanxun, QIN Lang
2023, 40 (5):  61.  doi: 10.3969/j.issn.2095-1736.2023.05.061
Abstract ( 38 )   PDF  
In order to study the ecological niche and interspecies association of dominant phytoplankton species after impounding of Wuxikou Water Conservancy Project in Changjiang River Basin, phytoplankton samples from nine sampling sites were studied in three different water situation periods in March(normal period), June(wet period) and November(dry period) 2021. The results showed that there were 27 dominant phytoplankton species in 4 phyla. Among them, Melosira granulata was the dominant species in all three situation periods, divided into widespread species. The appearance of Dinobryon divergens in November indicated that the water quality was οs type, showing a dynamic of poor nutrition, but the watershed in moderate condition. According to the niche breadth and niche overlap index of the dominant species, they were divided into three species with different resource utilization abilities. In addition, according to the results of statistical test, phytoplankton showed a significant positive correlation. The association was significant in 11 pairs. The significant rate of association was 6.18%. This may be due to the effect of water storage on phytoplankton species diversity, which makes the community structure in the early and middle stages of development.
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Immobilized surfaces demonstrate the adsorption properties of manganese peroxidase on methyl orange
WANG Yanjun, HUANG Jiayao, CHEN Shaohuang, ZHAO Pengfei
2023, 40 (5):  69.  doi: 10.3969/j.issn.2095-1736.2023.05.069
Abstract ( 49 )   PDF  
The yeast cell surface display vector pyd1 MnP was transformed into EBY100 by LiAC method. The MnP protein was successfully anchored on the surface of yeast cells by SDS-PAGE and immunofluorescence, and the transformant was induced to produce manganese peroxidase activity in vitro. It can provide excellent microbial resources for the degradation of azo dye wastewater. The enzyme activity was determined by 2,6-dimethoxyphenol (DMP) method, and the effects of different culture time, temperature, pH, carbon source, nitrogen source and concentration of carbon and nitrogen source on enzyme activity were determined. The enzyme was immobilized with sodium alginate. The optimal culture time and temperature of the immobilized transformant PM-2 were 60 h and 40 ℃. The optimal pH was 4.5, the optimal carbon source was galactose, the optimal nitrogen source was YNB, the carbon source concentration was 0.02 g/mL, and the nitrogen source concentration was 0.0072 g/mL. The adsorption performance of methyl orange simulated wastewater was studied by sodium alginate immobilization. The adsorption process accords with the quasi second order equation, and the fitting effect is better with Freundlich isotherm adsorption equation. The successfully anchored transformant PM-2 had the ability to produce manganese peroxidase, which can play a role in the biodegradation of azo dyes.
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Analysis and evaluation of agricultural non-point source pollution in Changlinhe Town
YAN Ting, LI Yucheng , SHI Yang, HU Yimin
2023, 40 (5):  77.  doi: 10.3969/j.issn.2095-1736.2023.05.077
Abstract ( 45 )   PDF  
The emission coefficient method was adopted to estimate the pollutant emissions in planting, livestock breeding, aquaculture industry and rural life of three small watersheds in Changlinhe Town. The pollution evaluation was carried out by equivalent pollution load method, combined with water quality monitoring results in-depth analysis of agricultural non-point source pollution. The results showed that the chemical oxygen demand (COD), total nitrogen (TN) and total phosphorus (TP) emissions in Changlinhe Town were 294.99, 56.41 and 6.41 t in 2021, respectively, and the COD emissions were the highest. The equivalent pollution load of the planting was the highest, accounting for 44.18% of the total emission standard, followed by rural life sources, accounting for 37.59%. The equivalent pollution load of TN accounted for 54.65% of the total emission standard, higher than that of TP and COD. Luodian, Luohong, Dongguang and Changlin community had a large equivalent pollution load, and the sum of the equal standard pollution load ratio of these four communities was 62.75%. Agricultural non-point source pollution was dominated by planting industry, in the future, pollution prevention and control should focus on planting industry and the main pollutant of TN should be controlled particularly, and the key control area is the Changlin River Basin.
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The role of prefrontal cortical GABAergic interneurons in the pathogenesis and treatment of depression
CHEN Zhibin, LIU Xiaoyu, ZHANG Mingkuan, GAO Qian, ZHANG Hao, LI Zifa, GENG Xiwen, WEI Sheng
2023, 40 (5):  83.  doi: 10.3969/j.issn.2095-1736.2023.05.083
Abstract ( 54 )   PDF  
This article reviewed the changes in morphology and function of the prefrontal cortex (PFC) and the alterations in the number and physiological function of GABAergic interneurons and their subtypes in depression. The impact of microstructural changes in different types of inhibitory neurons on the pathogenesis of depression, including their morphology and electrophysiological characteristics, was explored. The physiological and pathological changes in inhibitory neurons were discussed to elucidate their role in disinhibition of pyramidal neurons and the subsequent development of depression. The effects of antidepressant medications on GABAergic interneurons, including changes in their function and expression levels, were compared between preclinical and clinical studies. The involvement of GABAAand GABABreceptors as therapeutic targets in the treatment of depression was emphasized. Finally, the existing problems related to the involvement of GABAergic interneurons in the pathogenesis of depression in PFC were analyzed, and future research directions focusing on new receptors, targets, and pathways were proposed to provide insights into depression-related research and clinical treatment.
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Research progress of sucrose isomerase in food-safe bacteria #br#
CHEN Huiling, SHEN Haotian, JIANG Chen, LI Xianzhen, GUO Xiaoyu
2023, 40 (5):  88.  doi: 10.3969/j.issn.2095-1736.2023.05.088
Abstract ( 56 )   PDF  
The source, structure and reaction mechanism of sucrose isomerase in recent years were introduced. The research progress of heterologous expression of sucrose isomerase by food safety strains in recent ten years was summarized. The research pointed out the shortcomings of sucrose isomerase and prospected the future research direction from two aspects: the transformation of the expression host strain and the transformation of the encoding gene of sucrose isomerase.
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Research progress on immune regulation mechanism of animal antimicrobial peptides based on MAPK main signaling pathway
XU Chenlu, XIA Qiang
2023, 40 (5):  94.  doi: 10.3969/j.issn.2095-1736.2023.05.094
Abstract ( 64 )   PDF  
The latest research progress of animal antibacterial peptides in immunoregulation mechanisms was mainly introduced based on the MAPK signaling pathway in recent years. The research status of three MAPK subpathways, JNK, P38 MAPK and ERK, and the immunological regulation effects of animal antimicrobial peptides based on the MAPK pathway were summarized. The mechanisms by which antimicrobial peptides from mammals, birds, reptiles, amphibians, fish, insects and crustaceans through phosphorylation of MAPK pathway to involve in immune responses were also elucidated. The immunomodulatory mechanisms of various animal antimicrobial peptides based on the MAPK pathway were arranged and summarized, hoping to provide a literature reference for the comprehensive and in-depth research of the mechanism of antimicrobial peptides and the acting targets of antimicrobial peptides in the field of medicine.
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The current situation of population growth of crested ibis and its influencing factors
LIU Zunhai, ZHU Longbo, WANG Han, LEI Yinghu, ZHANG Junfeng, DONG Rong, SUN Chao
2023, 40 (5):  99.  doi: 10.3969/j.issn.2095-1736.2023.05.099
Abstract ( 65 )   PDF  
This article combined with the current conservation status quo and literature to summarize its living habits and population status. The main resistance in the development of the crested ibis population from both biological factors and environmental factors was analyzed. It specifically analyzed genetic factors, inbreeding, sex ratio, reintroduction, habitat and the impact of human activities on the development of the crested ibis population. The existing problems were pointed out at this stage and the suggestions were put forward to provide scientific guidance,such as continuing to adhere to the current reintroduction plan, while improving artificial breeding techniques and theoretical basis for the future protection of the crested ibis and population restoration and the stable growth of the crested ibis population.
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Research on the method of directly inducing callus from tobacco seeds
PENG Ye, XU Jiancheng, YU Lulu, HE Zhengquan, XU Fei
2023, 40 (5):  105.  doi: 10.3969/j.issn.2095-1736.2023.05.105
Abstract ( 55 )   PDF  
Tobacco seeds were used as explants to explore how to directly induce callus. The results showed that the callus induction of tobacco seeds was affected by different hormone ratios of 2,4-dichlorophe noxyacetic acid (2,4-D) and 6-benzylami nopurine (6-BA), and the combined use of hormones 2,4-D and 6-BA in the concentration range of 0.1-2.0 mg/L could induce callus, but different in size and quality. It was worth noting that when the seeds were cultured on the medium containing 1.0 mg/L 2,4-D + 1.0 mg/L 6-BA, callus was rapidly induced, but the callus mass was relatively loose. In comparison, the callus was more compact when the seeds were cultured on the medium containing 0.1 mg/L 2,4-D+1.0 mg/L 6-BA. Moreover, paraffin sections showed that the callus cells induced by the hormone combination of 1.0 mg/L 2,4-D+1.0 mg/L 6-BA were larger but irregularly arranged and most of them were non-embryonic cells. On the contrary, the callus cells induced by the hormone combination of 0.1 mg/L 2,4-D+1.0 mg/L 6-BA were small and regularly arranged, and most of them were embryonic cells. The results of this study provide new technical support for the induction of callus from tobacco seeds and facilitate the research on genetic transformation in tobacco.
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Efficient expression of human soluble transferrin receptor sTfR antigen and the preparation and application of its polyclonal antibody
LU Zhenhua, WU Zhenxing, YUAN Xinsong, WANG jian, WU Jianping
2023, 40 (5):  111.  doi: 10.3969/j.issn.2095-1736.2023.05.111
Abstract ( 57 )   PDF  
An efficient expression system of human soluble transferrin sTfR antigen was constructed by various strategies and the stable rabbit polyclonal antibody was obtained by immunization. High sensitivity in vitro detection kit was developed using sTfR antigen and antibody. A suitable expression system was chosen to express sTfR in human embryonic kidney cells 293 (HEK293), and the expression amount of sTfR was 0.52 mg/mL. To further improve the expression level of sTfR, the CMV promoter and the albumin signal peptide were added, leading to a significant increase in the amount of sTfR (0.86 mg/mL). High titer of rabbit sTfR polyclonal antibody was obtained using 0.3 mg sTfR antigen for the fifth immunization. When using sTfR kit to analysis human serum samples, the kit developed in this work was credible and stable. The development of sTfR kit and raw materials had significantly reduced the production cost. This work also offered an efficient approach for the development of other in vitro diagnostic reagents.
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Teaching reform and practice of veterinary microbiology experiment in application-oriented university
XIA Lunbin, CHEN Cunwu, ZHANG Xia, BI Shaoshuai, CHEN Naifu
2023, 40 (5):  116.  doi: 10.3969/j.issn.2095-1736.2023.05.116
Abstract ( 61 )   PDF  
Based on the existing deficiencies in the experimental teaching of veterinary microbiology and the research results of the job skills requirements of employers, the course team carried out teaching reform of veterinary microbiology experiments. First of all, by adjusting and optimizing the experiment content, it not only strengthened students’ grasp of basic experimental skills, but also paid more attention on the cultivation of comprehensive application ability. Secondly, the original single indoctrination teaching method was changed, and the internet mobile terminal and rain classroom auxiliary teaching platform were used to realize integrated teaching methods such as online and offline mixed classroom and flipped classroom. Furthermore, the evaluation method was optimized and the multi-element evaluation system was constructed to transform students from passive participants into the leader of the experiment. After more than 3 years of teaching practice, compared with that of the previous experimental teaching, the students’ initiative of participating in experiments and learning interest have been improved, the effect of learning was significant, improving their application and innovation abilities.
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Exploration and practice of opening and sharing platform for large-scale instruments training system
LI Mengyu, Li Bingyao, FAN Tao, CHEN Kai, YAN Xingrong
2023, 40 (5):  122.  doi: 10.3969/j.issn.2095-1736.2023.05.122
Abstract ( 57 )   PDF  
In view of the main problems existing in the traditional training mode, we have established systematic training methods, including developing a platform management system, raising awareness about instruments training, improving the skills of laboratory technicians, building a communication platform, a hybrid online and offline training system, students classification and personalized training, and constructing management team. The practice has shown that the comprehensive training system can enhance the skills and quality of users, which is conducive to increase the opening and sharing of large-scale instruments, to improve their utilization efficiency and to promote the output of scientific research results.
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Integrating theory and practice, creating a three-dimensional class of “teaching, learning, doing, thinking and using”
LI Xiaoyan, WANG Pengchao, BI Bing
2023, 40 (5):  126.  doi: 10.3969/j.issn.2095-1736.2023.05.126
Abstract ( 70 )   PDF  
Biochemical analysis technology explored on “the integration of theory and practice, and three-dimensional teaching model incorporating teaching, learning, doing, thinking and using” based on the student-centered concept. Theoretical teaching focused on optimizing the design of teaching content and selecting teaching methods to help students understand difficult points in the course more easily with the help of abundant online teaching resources. During practical teaching, zero-based experimental operations and teaching blank space design has been used in the first class to strengthen the foundation of experiments, to enhance operational skills, to inspire investigative spirit, and to promote the return of the student’s dominance in learning. By expanding the second classroom, we extended the knowledge from the first classroom and elevated students’ abilities, cultivated their advanced abilities such as exploration and creativity, and achieved deep learning. Practice showed that the three-dimensional teaching model promoted the interconnection and integration of the first and second classrooms in terms of form and content, and effectively created a new way to curriculum and teaching reform. It has significantly improved students’ ability to independently conduct scientific experiments and laid a solid foundation for their future independent scientific research.
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