Table of Content

18 December 2021, Volume 38 Issue 6 Previous Issue   

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Molecular mechanisms of plant in response to abiotic stress CHEN Keqi, DENG Xingguang, LIN Honghui 2021, 38 (6):  1.  doi: 10.3969/j.issn.2095-1736.2021.06.001 Abstract ( 3259 )   PDF   Save In this review, the molecular mechanisms by which plants respond to extreme temperature, drought, flooding and salt stress, through an overview of existing studies were summarized. Combining the effects of various stress on plant growth and development, the signal perception mode and signal transduction pathways were introduced. Meanwhile, the regulatory mechanisms of critical transcription factors in regulating gene expression and important secondary metabolites in coordinating stress tolerance were outlined. Finally, an outlook on future research was prospected focusing on the regulatory mechanism of mult-signaling network modules and sustainable development of agriculture. Related Articles | Metrics

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Cloning, bioinformatics analysis, subcellular localization and expressionanalysis of TaZFP33 gene from Triticum aestivum WU Qun, LI Yongliang, ZOU Xiaoxiao, SUN Aolong, LIN Xiaoxia, ZHOU Ping, GUO Xinhong 2021, 38 (6):  9.  doi: 10.3969/j.issn.2095-1736.2021.06.009 Abstract ( 2063 )   PDF   Save C2H2 zinc finger protein, as one of the largest transcription factor family in plants, plays an important role in plant growth and stress. In this study, the zinc finger protein TaZFP33 gene in the spring wheat variety Fielder（Triticum aestivum L. cv. Fielder） was cloned. The TaZFP33 gene coding region was 510 bp in length and encoded 169 amino acids. Bioinformatics analysis showed that the molecular weight of TaZFP33 protein was 17.5 ku, and the isoelectric point was 6.43, which belonged to the Q-type zinc finger protein and was an unstable hydrophobic protein. Evolutionary analysis and promoter analysis indicated that the gene may respond to ABA-induced stress regulation. Transient expression analysis of transformed tobacco showed that TaZFP33 was localized on the nucleus. Real-time fluorescence quantitative analysis of tissue-specific expression showed that TaZFP33 gene was expressed in stems, leaves, roots and embryos, and the highest expression level was in embryos. After seedlings were treated with different concentrations of ABA, PEG6000 and NaCl, the results showed that TaZFP33 gene responded to above-mentioned stresses. This research laid a foundation for further exploration on the biological function of the TaZFP33 gene, and also provided candidate genes for the study of stress resistance in Triticum aestivum. Related Articles | Metrics

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Codon usage bias and evolution analysis of the LaAP2 genein Lepidium apetalum Willd. ZHOU Qian, WANG Yuzhou, CHEN Yun, KAIDIRIYE·Yusupul, LIU Fei, ZHAO Huixin 2021, 38 (6):  15.  doi: 10.3969/j.issn.2095-1736.2021.06.015 Abstract ( 1889 )   PDF   Save To understand the codon bias of LaAP2 in Lepidium apetalum, LaAP2 gene codon bias was analyzed by Codon W, and EMBOSS programs was used to analyze the codon usage bias of the Lepidium apetalum LaAP2 gene. Correlation analysis was conducted to compare the bias of the Lepidium apetalum LaAP2 gene in model organisms for which relatively mature genetic transformation systems are available. Neutral drawing, ENc plots and parity rule 2 polt were subsequently used for the exploration of possible factors that affect the formation of the bias. The results showed that the ENc, CAI, and GC content of LaAP2 were respectively 48-65, 0-214, and 42-69%, indicating that the codon bias level of LaAP2 in Lepidium apetalum was low and biased toward the synonymous codons with A or T. According to the RSCU value, 11 codons showed high-frequency. Comparison of the codon usage preference of homolongous genes with other crops suggested that there was a certain difference in codon usage preference among AP2 genes in different species. The RSCU cluster analysis showed that the preference of LaAP2 gene was the closest to that of brassicaceae burnett in dicotyledonous plants. The codon base composition and correlation analysis showed that the codon bias of the Lepidium apetalum LaAP2 gene had many factors such as mutation and selective pressure playing an important role in shaping codon usage bias. Based on the analysis of the frequency of codon usage across species, the prokaryote E. coli expression system was found to be the most suitable for use with the LaAP2 gene, and the model plant Arabidopsis was the most ideal genetic transformation receptor of the LaAP2 gene. This study provided important reference information for further studies on the heterologous sexpression and the function of the LaAP2 gene in Lepidium apetalum. Related Articles | Metrics

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Prokaryotic expression and purification of mouse CCL2 protein MA Zhenling, WANG Lei, QIAO Liuhui, LIU Wei 2021, 38 (6):  20.  doi: 10.3969/j.issn.2095-1736.2021.06.020 Abstract ( 2174 )   PDF   Save his work aimed to construct a prokaryotic expression for mouse gene CCL2, express and purify recombinant CCL2 protein, and detect the adhesion of leukemia cells. The mouse gene CCL2 was amplified by PCR and cloned into the prokaryotic expression vector pGEX-4T-1 by ligation-independent cloning. The recombinant plasmid pGEX-4T-CCL2 was transformed into Escherichia coli BL21（DE3）and the recombinant protein was purified by affinity chromatography. The purified protein was identified by SDS-PAGE and Western Blot. Finally, the effect of CCL2 on C1498 cell adhesion and migration was detected. Results showed that the recombinant vector pGEX-4T-CCL2 was successfully constructed, and GST-CCL2 was successfully induced by 0.1 mmol/L IPTG at 20 ℃ for 6 h; recombinant protein with biological function was prepared, and CCl2 could promote leukemia cell adhesion and migration. This work would provide a foundation for further study of CCL2 in leukemia. Related Articles | Metrics

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Effects of fusion between breast cancer cells and macrophages on PI3K/AKT and MAPK/ERK signaling pathways ZHANG Lina, ZHANG Didi, ZHAO Lei, GU Yuxuan, SUN Xiaolin 2021, 38 (6):  25.  doi: 10.3969/j.issn.2095-1736.2021.06.025 Abstract ( 1843 )   PDF   Save In order to reveal the molecular mechanisms between cell fusion and tumor metastasis, we had successfully conducted a fusion model between breast cancer cells and macrophages. Previous studies found that the fusion hybrids exhibited significantly enhancing proliferation and metastasis abilities.Therefore, we next focused on the effects of cell fusion between breast cancer cells and macrophages on PI3K/AKT and MAPK/ERK signaling pathways closely related to tumor cell proliferation and metastasis. Western Blot results showed that the expression of the key marker proteins of PI3K/AKT and MAPK/ERK signaling pathways including p-PI3K, p-AKT and p-ERK1/2 was significantly up-regulated in the fused cells. After PI3K/AKT and MAPK/ERK signaling pathways being blocked by LY294002 and PD98059 inhibitors, the expression of p-AKT and p-ERK1/2 in the fused cells was obviously reduced, and the proliferation, migration and invasion abilities of the fused cells were inhibited. These results suggested that cell fusion between breast cancer cells and macrophages might promote the proliferation and metastasis of breast cancer by activating PI3K/AKT and MAPK/ERK signaling pathways. Our data also could provide new ideas for further understanding the mechanisms of cell fusion in tumorigenesis and metastasis. Related Articles | Metrics

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Coenzyme specific modification of NADPH-dependent glutamatedehydrogenase LU Libing, ZHOU Haisheng, WU Jianping, YANG Lirong 2021, 38 (6):  31.  doi: 10.3969/j.issn.2095-1736.2021.06.031 Abstract ( 2518 )   PDF   Save Based on structural information and conservative sequence alignment, this study modified the NADPH-specific glutamate dehydrogenase, from Pseudomonas putida, to NADH-dependent form in the future through semi-rational design and high-throughput screening. Compared that with the original PpGDH, the preference of the mutant PpGDH-D264V to NADH was increased by 1 607 times（Ratio of kcat/Km）, and it had the catalytic activity of various keto acids, which could be used to prepare various non-protein amino acids. The enzymic properties of PpGDH-D264V were characterized. The optimal temperature was 45 ℃, the optimal pH was 8.0, the temperature of T10 min1/2 reached 58.1 ℃, and the PpGDH-D264V had good stability under neutral and slightly alkaline environment, which laid a foundation for subsequent application. Related Articles | Metrics

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Study on the Cr(VI) reduction properties of Alicycliphilus denitrificans Ylb10 ZUO Qun, LYU Yucai, SONG Tingwei, GUO Jinling, REN Liwei, GONG Dachun 2021, 38 (6):  36.  doi: 10.3969/j.issn.2095-1736.2021.06.036 Abstract ( 2768 )   PDF   Save From the bottom mud of Qiusuo Creek, a strain Ylb10 with high efficiency in reducing Cr(VI) was successfully isolated and obtained. The Ylb10 strain was identified by physiological and biochemical methods and 16S rDNA sequencing method, and the Cr(VI) was determined by diphenylcarbazide spectrophotometry to investigate its reduction characteristics. The results showed that Ylb10 was Gram-negative, and the 16S rDNA similarity rate with Alicycliphilus denitrificans was 98.07%. Ylb10 could make good use of α-hydroxy-butyric acid, L-lactic acid, propionic acid, acetic acid, L-aspartic acid, L-glutamic acid, L-alanine, L-erine and other substrates.Compared that with the shaker culture conditions, Cr(VI) could be better reduced during standing culture, and the optimal pH of Cr(VI) reduction was 8-9. When the concentration of Cr(VI) was lower than 200 mg/L, the strain could reduce all Cr(VI), of which, at 50 mg/L Cr(VI) concentration, it had a reduction rate of 96.45% in 18 hours; at 100 mg/L Cr(VI ), it had the reduction rate of 93-83% in 24 hours, and the reduction efficiency at 200 mg/L Cr(VI) concentration in 60 hours was 99.06%. It could be concluded that Alicycliphilus denitrificans Ylb10 was a high-efficiency hexavalent chromium reducing bacteria, and It had high application potential in the treatment of chromium-contaminated water quality and soil. Related Articles | Metrics

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Seasonal variations of thermoregulation and thermal neutral  zone shift in Eothenomys olitor YANG Yazuo, JIA Ting, ZHANG Hao, WANG Zhengkun, ZHU Wanglong 2021, 38 (6):  41.  doi: 10.3969/j.issn.2095-1736.2021.06.041 Abstract ( 1181 )   PDF   Save In order to explore the physiological adaptation characteristics of Eothenomys olitor in Zhaotong area, body temperature, resting metabolic rate (RMR) and thermal conductance (C) were measured in the range of 5℃-30℃ in summer and winter, respectively. The results showed that there were seasonal differences in body temperature, RMR and C value in E. olitor. Body temperature in summer was higher than that in winter, and the average body temperature was 36.17℃±0.52℃ and 35.55℃±0.46℃, respectively. The thermal neutral zone (TNZ) showed seasonal differences. TNZ was 20℃-27.5℃ in summer and 15℃-25℃ in winter respectively, which showed that the lower limit critical point in winter was lower than that in summer. In TNZ, RMR in summer (2.76±0.19) mL O2/(g·h) was lower than that in winter (4.78±0.22) mL O2/(g·h). In summer and winter, C value in E. olitor was stable in TNZ, and the mean values were （0.18±0.02） and （0.31±0.04）mL O2/(g·h·℃), respectively. The above results suggested E. olitor adapted to the climate characteristics of Zhaotong area, which were large temperature difference between winter and summer and low annual extreme temperature by regulating body temperature, RMR and C value. Related Articles | Metrics

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Effects of the medicines for preventing and controlling Sclerotinia sclerotiorum on growth and development of silkworm WU Fan, LI Dechen, HE Zhen, CHEN Dengsong 2021, 38 (6):  45.  doi: 10.3969/j.issn.2095-1736.2021.06.045 Abstract ( 332 )   PDF   Save This study explored the effects of three mulberry sclerotium control drugs on silkworm growth and development. The second-instar (2nd) silkworms were fed with carbendazim, procymidone and thiophanate-methyl until the cocoon stage, respectively, taken fresh water as a control. The effects on the silkworm growth and development were measured by the length of larval stage, the body weight of the fifth instar, and the cocoon quality. Also the activity of antioxidant enzymes in the midgut of the fifth instar larvae on the fourth day was measured by CAT and POD activity kits. The larvae fed with thiophanate-methyl could not complete the generational development, which died at the 2nd instar. The silkworms fed with carbendazim and procymidone could normally develop into cocoon stage. However, compared that with the control group, the larval age of silkworms fed with carbendazim had shortened, while the age of silkworms with procymidone had increased. The weight of silkworms fed with carbendazim and procymidone became lighter, which suggested that these two powders had a negative effect on the cocoon performance of silkworms. The activities of CAT and POD enzymes in the midgut of silkworms had increased. Therefore, for the control of mulberry sclerotium, the usage of thiophanate-methyl should be prohibited, while carbendazim and procymidone might be used. Related Articles | Metrics

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Intestinal parasite diversity of the wintering lesser white-fronted goose (Anser erythropus) and sympatric poultry ducks in Shengjin Lake, China GONG Zhizhong, LI Qingyue, WANG Chong, XU Caoling, LIU Gang 2021, 38 (6):  49.  doi: 10.3969/j.issn.2095-1736.2021.06.049 Abstract ( 313 )   PDF   Save From November 2017 to April 2018, 350 Anser erythropus faecal samples and 350 sympatric poultry duck faece samples were collected and detected with saturated brine floating and centrifugal sedimentation methods in Shengjin Lake. Parasite community structure and prevalence 13 parasites were observed from 350 wintering Anser erythropus faecal samples, about 67.4% showed parasitic infection. Eleven parasites were observed from 350 poultry duck faecal samples with the infection rate of 70.1%, and considerable differences were found in the infection intensity among parasite species. The most abundant species were coccidian oocysts, followed by Nematode and Trematode, while Cestode made a rare appearance in Anser erythropus and poultry duck. Most positive samples contained one or two species of parasites, samples with a single species of parasites were more common than those containing two or more. Examination of Anser erythropus and the poultry duck samples collected from Shengjin Lake revealed that some parasites were recorded in other waterbirds and poultry birds. The similarity coefficient between Anser erythropus population and the poultry duck population was 0.43. It might be the similar diets, high population densities and niche overlap leading to the similar parasite community of Anser erythropus population and the poultry duck in Shengjin Lake, this similarity coefficient might pose potential hazards to cross transmission between Anser erythropus population and the poultry duck population. The similarity of parasitic community structures in the wintering Anser erythropus and poultry bird populations, suggested that parasitic epidemic was more sensitive in the overwintering period in the Shengjin Lake, and might also cause human disease, even cause potential public health risks. Related Articles | Metrics

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Study on genetic diversity of tea tree from Fujian Province based on matK partial sequence NIE Chuanpeng, CHEN Dandan, LI Yanyan 2021, 38 (6):  54.  doi: 10.3969/j.issn.2095-1736.2021.06.054 Abstract ( 248 )   PDF   Save In order to study the genetic diversity of Fujian tea trees, the genetic structure and genetic differentiation of 51 cultivars of Fujian tea trees from 5 regions were analyzed. The results showed that the length of matK sequence of tea plants in Fujian was 833 bp, with 26 polymorphic sites (S) and 7 haplotypes (Hap1-Hap7), showing high haplotype diversity (0.525) and low nuclear glycoside diversity (0.001 89). The haploid phylogenetic tree formed 4 branches, and the self-expanding support rate was greater than 60%. The genetic distance between different tea populations was between 0.000 3 and 0.003 3, and the genetic distance within the population was between 0.000 0 and 0.003 7. The differentiation coefficient of Fujian tea tree population was Fst=-0.016 22-0.500 00 and gene flow was Nm=-15.663 07-15.210 73. The genetic diversity of 51 Fujian tea trees was low, and there was a certain degree of genetic differentiation between different groups. Neutral tests Tajima’s D, Fu and Li’D* and F* were significantly negative values, and the mismatch distribution curve was a unimodal pattern, indicating that expansion events occurred among the tea tree populations. The results of this study provided a reference for further development and utilization of this resource. Related Articles | Metrics

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A cladistic analysis of Rhizocarpon based on morphological characteristics HANAT Gulina, LI Zuosen, TUMUR Anwar 2021, 38 (6):  59.  doi: 10.3969/j.issn.22095-1736.2021.06.059 Abstract ( 279 )   PDF   Save The aim of our study was to analyze morphological and anatomical characters of the 22 species belonging to the lichen genus Rhizocarpon, found in Xinjiang, China, to provide a morphological basis for the interspecific classification. Twentythree morphological characters of 22 species of Rhizocarpon were coded. Catolechia wahlenbergii and Epilichen scabrosos, belonging to the genus Epilichen and Catolechia in the family Rhizocarponaceae, were designated as outgroup. Their morphological characteristic matrix was analyzed by Maximum Parsimony(MP) method using PAUP, compared with the results of molecular phylogeny based on ITS sequence. After parsimonious analysis, the equally most parsimonious cladograms were obtained, and its strict consensus tree showed that the species of genus Rhizocarpon were divided into two branches according to the type of ascospores, which were composed of species with 1septate ascospores and the species with (sub)muriform ascospores, respectively. R. parvum and R. pusillum, R. geographicum and R. atroflavescens as well as R. macrosporum, R. saurinum and R. furax were gathered in one branch of the phylogenetic trees, respectively.The results of this study were generally consistent with that of molecular phylogenetic analysis. The evolutionary position of some species based on the results of molecular phylogeny was different from that of the phylogenetic trees based on morphological characteristic. From the results of cladistic studies, the type of ascospores was an important taxonomic characters of the genus Rhizocarpon, and the present cladistic study would provide a morphological basis for the evolutionary position of the genus Rhizocarpon. Related Articles | Metrics

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Fourier infrared spectrum analysis of Dendrobium nobile from Qiyun Mountain, Anhui Province TIAN Shengni, LI Yanan, HU Yixuan, LI Chun 2021, 38 (6):  65.  doi: 10.3969/j.issn. 2095-1736.2021.06.065 Abstract ( 318 )   PDF   Save The infrared spectra of roots, stems and leaves of Dendrobium nobile from Qiyun Mountain, Anhui Province, Dendrobium officinale from Lu’an and Wenzhou areas were determined by Fourier transform infrared spectroscopy in this paper, which aimed to explore the infrared spectral characteristics of Dendrobium nobile from Qiyun Mountain and the differences of chemical components among them. It was found that the infrared absorption peaks of roots, stems and leaves of Dendrobium from three areas were similar in shape and position, mainly distributed around 3 420, 2 920, 1 730, 1 240, 1 320 and 1 150 cm-1, but there were some differences in the intensity of each absorption peak. Their chemical compositions were basically the same, including polysaccharide, protein, tertiary amide, lignin, etc, but their contents were different. In the fingerprint region of 900500 cm-1, the similarity between the stem and leaf of Dendrobium nobile from Qiyun Mountain was higher than that of root. In the range of 1 500500 cm-1, there were significant differences in infrared spectrum peak shape and peak intensity of roots, stems and leaves of Dendrobium from three areas. The results showed that Fourier transform infrared spectroscopy could effectively identify the similarities and differences of chemical components among different Dendrobium germplasm resources. The leaves and stems of Dendrobium nobile from Qiyun Mountain, Anhui Province have similar infrared spectra, and the leaves of Dendrobium nobile from Qiyun Mountain can be used as a medicinal substitute for its stems. Related Articles | Metrics

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Screening and performance of high-efficiency root-knot nematodebiocontrol fungi LI Lei, ZHAO Junjie, LIU Yingying, ZHEN Jing, LI Liangliang, CHEN Guocan, MU Qi, WANG Jiwen 2021, 38 (6):  70.  doi: 10.3969/j.issn.2095-1736.2021.06.070 Abstract ( 282 )   PDF   Save In order to obtain strains with the effect of preventing and controlling root-knot nematodes, the chitin flat transparent ring primary screening and the in vitro antagonistic test rescreening methods were used to obtain a fungal strain SFC-3 with biocontrol potential. Through observation of morphological characteristics and analysis and identification of transcribed spacer sequences (ITS) in fungi, the strain SFC-3 was identified as Trichoderma asperellum and its chitinase activity was 2.07 U/mL. The root-knot nematode egg parasitic experiment and the second-instar larval lethality experiment showed that the strain SFC-3 had the highest parasitic rate of 78.86%, which was higher than SFC-1, SFC-22 and other 9 strains. At the 7th day, the relative inhibition rate of egg incubation of root-knot nematodes reached 96.51%, and the corrected mortality rate of second-instar larvae reached 93.37%. In this study, by screening anti-knot nematode strains and studying their biocontrol effects, it was confirmed that the strain SFC-3 had better anti-root knot nematode performance. Related Articles | Metrics

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The response of soil microbial diversity to the difference of Ndeposition concentration in alpine wetland XU Runhong, TAN Mei, ZHU Jinfu, LIU Zehua 2021, 38 (6):  75.  doi: 10.3969/j.issn.2095-1736.2021.06.075 Abstract ( 271 )   PDF   Save In order to understand the response of soil microbial diversity to nitrogen deposition in alpine wetland under the background of nitrogen deposition, four levels (0, 2, 5 and 10 g/m2) were used to simulate nitrogen addition. The diversity and community structure of soil bacteria and fungi were analyzed by high throughput technique. The results showed that the dominant flora of bacteria were Acidobacteria and Actinobacteria of Proteobacteria and that of fungi were Ascomycota Basidiomycota and Mortierellomycota. Nitrogen treatment significantly changed the relative abundance of bacteria and fungi. NMDS analysis showed that the community structure of soil bacteria and fungi changed significantly under nitrogen treatment (P＜0.01). Cluster analysis showed that the difference of soil microbial species composition increased with the increase of nitrogen application concentration. With the change of nitrogen application concentration, the diversity index of microorganisms changed significantly in the 015 cm soil layer, but not significantly in the 1530 cm soil layer. The response mechanism of bacteria and fungi to N deposition was different, the bacterial diversity showed an upward trend, while the fungal diversity showed a downward trend. It can be seen that N deposition has an impact on soil microbial structure and a certain difference on bacterial and fungal diversity, which can provide a basis for wetland management under the background of N deposition. Related Articles | Metrics

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Correlation analysis between gut microbiota and biochemical indicators of cirrhosis LI Lili , XIE Ruiqi, WANG Li , LIU Youde, DENG Lixia, ZOU Zhiqiang, QIN Song 2021, 38 (6):  82.  doi: 10.3969/j.issn.2095-1736.2021.06.082 Abstract ( 1054 )   PDF   Save In order to characterize the gut microbiota of patients with cirrhosis and reveal the association between gut microbiota and biochemical indicators, gut microbiota and biochemical indicators of 12 cirrhosis patients (CIR group) and 11 healthy volunteers (control, CN group) were analyzed. Compared with that of the CN group, the levels of INR, GGT, TBA and TBIL in the CIR group increased significantly, while the levels of ALB and WBC decreased significantly. Gut microbiota analysis indicated that the abundance of genus Prevotella and Ruminococcaceae UCG-014 increased significantly, while genus Blautia, Subdoligranulum and Ruminococcus 2 decreased significantly in the CIR group. At the species level, the abundance of Prevostella 9 and Ruminococcaceae UCG-014 increased significantly, while Ruminococcus sp. 5 1 39BFAA decreased significantly in the CIR group. The correlation analysis between biochemical indexes and intestinal flora showed that the abundance of Prevotella 9 genus had significantly positive correlation with INR, TBA and GGT levels, and negative correlation with ALB level. The abundance of Ruminococcaceae UCG-014 genus was positively correlated with INR and GGT levels. The abundance of Blautia genus demonstrated significantly positive association with ALB level, and negative association with INR, TBA, TBIL and GGT levels. The abundance of Subdoligranulum and Ruminococcus 2 genera exhibited significantly positive correlation with WBC level and negative correlation with TBA and INR levels. At the species level, the abundance of Ruminococcus sp.5 1 39BFAA showed significantly positive correlation with ALB level, and significantly negative correlation with INR, TBA, TBIL and GGT levels. In summary, cirrhosis led to abnormal biochemical indexes of liver and imbalance of intestinal flora. Prevotella genus, Blautia genus, Ruminococcaceae UCG-014 genus, Subdoligranulum genus and Ruminococcus sp.5 1 39BFAA in patients with cirrhosis were significantly different from the healthy group, and these bacteria were related to liver damage. The discovery of these characteristic bacteria and their relationship with biochemical indexes were expected to provide potential targets for precise treatment of cirrhosis. Related Articles | Metrics

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Proximity labeling technique and its application and research prospect in neurobiology FAN Lingling, SHEN Guomin 2021, 38 (6):  87.  doi: 10.3969/j.issn.2095-1736.2021.06.087 Abstract ( 801 )   PDF   Save The principle, application and common problems of peroxidase-based PL and biotin ligase-based PL are introduced. This review focuses on the current application of PL technique in the study of synaptic subregion molecular components in the field of neurobiology and its achievements, and analyzes the challenges faced in the application process, such as probe input and high background in living brain tissue. It is pointed out that this technique will provide an unprecedented opportunity to systematically identify the highly complex proteome of dynamic interaction and subcellular structure in neural tissue, and also provide an effective approach to search for key molecules in neurobiology. Related Articles | Metrics

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Humanized antibody transgenic animal platforms and its patent protections WU Meng, ZOU Xiangang, LIU Zuohua, YANG Songquan , GE Liangpeng 2021, 38 (6):  92.  doi: 10.3969/j.issn.2095-1736.2021.06.092 Abstract ( 280 )   PDF   Save This paper reviewed the evolution of therapeutic antibody drugs from mouse antibody, chimeric antibody, humanized antibody to full human antibody, as well as the development process of humanized antibody transgenic animals used for full human monoclonal antibody discovery. The characteristics and core patent protection of human antibody transgenic animal platform commonly used in the world were mainly summarized. Since human antibody transgenic animals use genetic engineering technology to replace animal immunoglobulin gene with human antibody immunoglobulin gene, it can directly express human antibody protein, so it has been widely used in the development of whole human antibody drugs. However, in order to solve the matching problem between the human immunoglobulin gene and animal B cell development, obtain the candidate antibodies with richer diversity and higher affinity, it is necessary to conduct more in-depth basic research on the mechanism of animal antibody production. At the same time, in the face of sudden biosafety incidents, how to rapidly and massively produce full human antibodies is also a key issue to be considered in the future cultivation of humanized antibody transgenic animals. Related Articles | Metrics

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Research progress on African swine fever virus and its vaccines HUANG Xia, KANG Xilong, MENG Chuang , GU Dan, JIAO Xin’an, PAN Zhiming 2021, 38 (6):  99.  doi: 10.3969/j.issn.2095-1736.2021.06.099 Abstract ( 353 )   PDF   Save Taking African swine fever virus (ASFV) as the research object, pathogenic characteristics of ASFV including virus structure, infection and immune mechanism, transmission route, and pathogenicity were summarized. Meanwhile, the research status of African swine fever (ASF) vaccines was analyzed: inactivated vaccines cannot provide protection; subunit vaccines, nucleic acid vaccines and viral vector vaccines usually provide partial protection; live attenuated vaccines can provide homologous protection and are most potential for clinical application. On this basis, it was found that the complexity of ASFV and the uncertainty of immunobiological characteristics affect the progress of vaccine research. And the challenges in development of ASF vaccines faced by China were put forward to providing a new understanding and approach for vaccine development. Related Articles | Metrics

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Research advance of plant metallothioneins in metal ion combination and detoxification ABLIMIT Mehriban, WANG Yan 2021, 38 (6):  104.  doi: 10.3969/j.issn.2095-1736.2021.06.104 Abstract ( 455 )   PDF   Save Plants induce the metallothioneins (MTs) to regulate metal homeostasis, buffer toxic heavy metals and remove cell-damaging reactive oxygen species (ROS) under metal stress. Building upon functions on plant metallothioneins, the functions and mechanisms of metal ion binding ability and ROS scavenging at home and abroad for the past few years were reviewed. The chemical basis of MTs ability to bind different metal ions based on the characteristics of ligand-to-metal charge transfer (LMCT) was summarized. MTs played important roles in metal homeostasis and heavy metal detoxification. Furthermore, the thiol groups in MTs could be oxidized. MTs interacted with the integral membrane proteins in the outer mitochondrial membrane, and indirectly regulated the activities of antioxidant enzymes or related genes expression to eliminate ROS in plants under heavy metal stress. Related Articles | Metrics

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fast and sensitive method for determination of non-specific nuclease activity GAO Ting, ZHU Yu, ZHAO Chen, GE Yan, LI Duanhua, LI Jinjun, WANG Lu 2021, 38 (6):  111.  doi: 10.3969/j.issn.2095-1736.2021.06.111 Abstract ( 334 )   PDF   Save A rapid, simple and high-throughput activity assay method of non-specific nuclease has been established, according to the fact that under the irradiation of ultraviolet light and through the combination of nucleic acids with DNA stain Gelred.The fluorescent complex was generated and then emited fluorescence. In the study, the change of optical density caused by the decrease of substrate DNA measured by a gel imager, therefore the enzyme activity was obtained. Key factors involved in the activity assay experiment were studied, as well as the linearity, accuracy, precision, limit of detection (LOD) and limit of quantification (LOQ) of the method were all verified. The results suggested that the optimal reaction time was 3 min, the linear range of enzyme activity was 4.17-9.01 U/μL, and the linear relationship, precision, accuracy and sensitivity met the evaluation requirements of methodology development. In conclusion, the method exhibited lower LOD and shorter reaction time compared with the UV absorption assay method, and it could be applied for multiple samples analysis. Related Articles | Metrics

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The reform of microbiology experiment examining under “3+1”teaching mode MA Hongmei, LI Zhen, MA Jun 2021, 38 (6):  116.  doi: 10.3969/j.issn.2095-1736.2021.06.116 Abstract ( 216 )   PDF   Save In order to make up for the disadvantages of traditional microbiology experiment assessment, verification and comprehensive experiments were no longer set up alone, just one large comprehensive experiment was designed with the condition of limited teaching hours, which was disintegrated into eight small experimental items with verification, synthesis and design. Under the “3+1” experimental teaching mode, each small experimental item was quantitatively scored according to the four steps of experimental preparation, intelligent guidance, experimental operation and summary innovation, the final experiment examination was no longer arranged, and the experimental scores of students were comprehensively determined according to the scores of each step of each small experiment item done in the “3+1” teaching mode. The degrees of the students’ learning and understanding for each small experimental item could be judged based on the analysis of each highscore segment score of each small experimental item. Compared with the traditional microbiology experiment teaching examining method, the examination of microbiology experiment under the mode of “3+1” teaching reform paid more attention to the whole process of each small experimental item to make a quantification, the examination standard was clear, the examination content was comprehensive as well as the offline assessment of the examination mode was combined with online, which reduced the probability of score teachers assess randomly; the probability of speculative score of students and effectively promoted the students to participate in the whole teaching of the experiment, and their practical ability and ability of innovative summary were both improved. It also enabled teachers to acquainte objectively with the effect of experimental teaching and learning of students, and find out the problems existing in experimental teaching exactly and improve them. Related Articles | Metrics

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Application of multiple teaching mode oriented by creation and innovation education in the practice teaching process of metabolic control fermentation LIU Huiyan, FANG Haitian, PAN Lin 2021, 38 (6):  119.  doi: 10.3969/j.issn.2095-1736.2021.06.119 Abstract ( 200 )   PDF   Save The traditional curriculum practice teaching mode is teacher centered, and most of the practice sessions are mainly based on verification and visiting, students’ experience and driving force for understanding and mastering knowledge points, operation points and experimental methods were insufficient. And, it cannot meet the training requirements of students with innovative thinking and ability. Therefore, the practice teaching of this course has been reformed. In the teaching process, the multi-level practice teaching method oriented by innovation and entrepreneurship was adopted, which embodied the concept of student-centered cultivation and integrated innovation and entrepreneurship education into professional practice teaching. We realized the organic integration of the first classroom and the second classroom, improved the students’ learning effect, and drove the students’ creation and innovation passion using this teaching mode. Related Articles | Metrics

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Exploration and practice of online and offline mixed teaching mode of biochemistry experiment ZHANG Shaobin 2021, 38 (6):  123.  doi: 0.3969/j.issn.2095-1736.2021.06.123 Abstract ( 139 )   PDF   Save In order to meet the needs of cultivating high-quality talents in biological science major, based on modern information technology, the independent campus SPOC of biochemistry experiment was built on the public MOOC platform, and the online and offline mixed teaching modes were explored and practiced. The teaching process of each experimental project was divided into four main stages: pre-class preparation, online teaching, offline teaching and after-class summary. Online teaching mainly completed the teaching contents of experimental principle, reagent preparation, instrument use and demonstration of experimental operation process, while offline teaching was mainly based on students’ hands-on operation. Online teaching was well prepared for offline teaching, which not only improved the efficiency of offline teaching, broadens students’ vision by experimental items of expanded, improved students’ comprehensive quality through ideological and political education, but also improved the objectivity and accuracy of curriculum assessment, increased the degree of grade differentiation through the combination of online assessment and offline assessment, improved students’ interest and autonomous learning ability, realized the cooperative cultivation of knowledge, quality and ability, and provided a reference and guidance for the reform of online and offline mixed teaching mode in other experimental courses. Related Articles | Metrics

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Teaching practice of bioengineering comprehensive experiment in local universities ZHANG Xuze, WANG Jiuli, WANG Kezhou, WANG Xueren, SHEN Yingfang 2021, 38 (6):  127.  doi: 10.3969/j.issn.2095-1736.2021.06.127 Abstract ( 236 )   PDF   Save In order to improve the education level of bioengineering major in local universities, based on the purpose and requirements of bioengineering comprehensive experiment teaching and combining with local characteristics, a PBL model of bioengineering comprehensive experiment teaching was constructed. This model aimed to improve students’ practical innovation ability. The experimental design was completed by a project team composed of students, which associated the course content with the product production practice, and completed the whole process of the experiment with project management. The teaching reform and practice were carried out from the aspects of course orientation, teaching content setting and teaching means innovation. Through the integration of discipline teaching and research ability cultivation, the reform mode of bioengineering practice system from basic to comprehensive practice was embodied. Related Articles | Metrics