18 April 2024, Volume 41 Issue 2 Previous Issue   
Research progress on carbon reduction potential of agriculture in China
ZHOU Xinyan, ZHENG Huabao
2024, 41 (2):  1.  doi: 10.3969/j.issn.2095-1736.2024.02.001
Abstract ( 21 )   PDF  
This review provided an overview of the main characteristics and types of agricultural carbon emissions in China. It also identified potential areas for carbon reduction in agriculture, named farmland utilization, livestock and poultry breeding, agricultural waste disposal, and land use management. Reasonable agronomic management and economic policy measures can effectively reduce carbon emissions from agricultural land use. Measures such as reducing agricultural carbon emissions at the source, implementing process control, and employing end-of-pipe treatment techniques are effective in mitigating carbon emissions from livestock and poultry farming. Additionally, optimizing manure management and agricultural straw utilization can minimize carbon emissions from the disposal of agricultural waste. Furthermore, adopting an optimized structure for land resource utilization, implementing intensive land use practices, and constructing high-standard farmland are crucial measures for carbon reduction in land use management. This article serves as a valuable reference for enhancing the potential of carbon reduction in China’s agriculture.
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creening and heterologous expression of ω-transaminase for the preparation of S-methoxyisopropylamine
ZHANG Tao, ZHOU Haisheng, XU Jiaqi, YANG Lirong
2024, 41 (2):  8.  doi: 10.3969/j.issn.2095-1736.2024.02.008
Abstract ( 14 )   PDF  
An enzyme library containing 36 ω-transaminase was constructed according to the evolutionary classification of the aminotransferase family and substrate specificity. Genes from the library were cloned into E.coli BL21 (DE3) for heterologous recombinant expression of the enzymes. The enzyme expression levels were analyzed and corresponding enzyme activity and enantiomeric selectivity were determined. Through screening, the best ω-aminotransferase was found to be the one from Bacillus megaterium (BmeTA), which exhibited a crude enzyme activity of 2.0 U/mL and a pure enzyme activity of 9.5 U/mg protein. Enzymology characterization showed that the optimal temperature of BmeTA was 35 ℃, and the optimal pH was 8.0. Based on these, the catalytic reaction process conditions were further optimized, and 450 mmol/L of S-methoxyisopropylamine was obtained after an 18 h catalytic reaction under the conditions of 20 g/L crude enzyme and 0.5 mmol/L coenzyme dosage, and 1.4 amino donor/amino acceptor ratio, reaching a 90% conversion rate of substrate. The investigation of this study laid the foundation for the industrialization of biocatalytic preparation of S-methoxyisopropylamine.
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Recombinant expression and biochemical characterization of the raw starch digesting α-amylase Amy486
CHEN Xin, GAO Huanhuan, WANG Bin, ZHANG Xuecheng, FANG Zemin, XIAO Yazhong, FANG Wei
2024, 41 (2):  16.  doi: 10.3969/j.issn.2095-1736.2024.02.016
Abstract ( 16 )   PDF  
To characterize the novel raw starch digesting α-amylase Amy486,amy486was cloned from the marine bacterium Exiguobacterium sp. J84 and expressed heterologously. After purified by Ni2+-NTA affinity chromatography column, the catalytic property, halophilic property and Ca2+-dependence of Amy486 were analyzed. The optimum pH of Amy486 was 7.5 and it maintained above 40% residual activity in the pH range of 6.5-8.5. The optimum temperature was 35 ℃ and Amy486 was more stable at lower temperature, with a half-life of about 100 h at 30 ℃. With the addition of 1.5 mol/L Na2SO4, the specific activity toward raw rice starch reached 2209 U/mg. In the presence of 1.0 mol/L Na2SO4, Amy486 maintained more than 60% relative activity at 35 ℃. The enzymatic activity can be enhanced up to 110% in the presence of 2.5 mmol/L CaCl2, and the activity was inhibited with the addition of more than 5 mmol/L CaCl2. K302 was determined as the binding site of calcium ion. The mutant K302E exhibited enhanced binding ability to calcium ion. Amy486 and K302E are halophilic raw starch digesting α-amylases and low dependence on calcium ions, which possess potential application in hydrolysis of starch in high salt environment.
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Reassembly between two distinct G-quadruplexes
BI Xinmeng, FU Wenqiang, ZHANG Na, WANG Tao
2024, 41 (2):  25.  doi: 10.3969/j.issn.2095-1736.2024.02.025
Abstract ( 10 )   PDF  
Here, mainly through solution NMR experiments, we found that the mutant sequence of Thrombin Binding Aptamer (TBA-M) and human telomeric sequence htel3 self-fold into G-quadruplexes in sodium solution, respectively. Appealingly, these two well pre-folded G-quadruplexes can further reassemble into a new heteromolecular G-quadruplex complex (TBA-M/htel3) spontaneously through DNA strand displacement. The stoichiometric ratio between TBA-M and htel3 in the complex is identified as 1∶1. The last three guanosines (G14G15G16) from the 3′-end of TBA-M are involved in the formation of the G-quadruplex core of TBA-M/htel3 complex via Hoogsteen base pairing. In this paper, we reveal for the first time that two well pre-folded G-quadruplexes enable a further Hoogsteen pairing-based DNA strand displacement, in which the reaction process and molecular mechanism are also investigated primarily. Our finding expands the understanding of the interacting pattern and recognition mechanism between differently folded structures of nucleic acids.
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The molecular mechanism of RPS6 subunit peptide segment inhibiting S180 tumor cells
PU Dihong, YE Ziyu, ZHOU Liqian, LIU Xinlan, LU Yan, HOU Yiling, DING Xiang
2024, 41 (2):  32.  doi: 10.3969/j.issn.2095-1736.2024.02.032
Abstract ( 6 )   PDF  
To explore the effect of ribosomal protein RPS6 subunit peptide on S180 tumor cell gene expression and the key molecule and signal pathway of inhibiting tumor cells, using S180 tumor bearing mice as models, S180 tumor bearing mice were treated with RPS6 subunit peptide, and S180 tumor cells were sequenced by Illumina to obtain differentially expressed genes, and these differentially expressed genes were analyzed by GO and KEGG. Gene expression results showed that RPS6 subunit peptide could reduce the expression of mt-Cytb,mt-Nd1andRpl13genes, and hinder the oxidative phosphorylation process of S180 tumor cells. The results of GO analysis showed that the key categories of RPS6 subunit peptide inhibiting tumor cells focused on the changes of plasma membrane and its outer components, and the regulation of immune response and the activation of immune cells. The differential gene results showed that RPS6 subunit peptide could enhance PRL/PRLR signal, up regulateUchl1gene expression and induce tumor cell cycle arrest. The results of KEGG pathway enrichment showed that the expression of perforin encoded byPrf1and granzyme B encoded by Gzmb, together with cytotoxicity mediated by natural killer cells, participated in S180 tumor cell apoptosis under the effect of RPS6 subunit peptide. Meanwhile, TNF-α expression was up-regulated in NF-κB signal pathway, in coordination with up-regulated IFN-γin cell-mediated cytotoxicity pathway also jointly inhibiting the proliferation of S180 tumor cells. RPS6 subunit peptide led to cell cycle arrest of S180 tumor cells, induced cytotoxicity mediated by natural killer cells and up regulation of NF-κB signal pathway led to S180 tumor cell apoptosis, which provided a theoretical basis for the application of RPS6 subunit peptide in antitumor research.
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Anti-inflammatory effect and mechanism of Pseurata H on inflammation model of LPS-induced RAW264.7 cells
YANG Dongjuan, ZHA Guangcai, ZOU Xianghui, WU Dantong, WU Qingting
2024, 41 (2):  39.  doi: 10.3969/j.issn.2095-1736.2024.02.039
Abstract ( 8 )   PDF  
To study the anti-inflammatory effect and mechanism of Pseurata H on peritoneal macrophages (RAW264.7 cells) inflammation models, RAW264.7 cells were stimulated with LPS to establish an inflammation model in vitro, and Pseurata H with the concentrations of 10, 20, and 40 μmol/L was used to interfere with this cell model. The cytotoxicity of Pseurata H in RAW264.7 cells was determined by CCK-8 colorimetric assay. Griess assay was employed to detect the content of NO in the supernatant. The release of IL-6 and TNF-α in the supernatant was assayed using ELISA. The protein expressions of COX-2, iNOS, p-NF-κB and the p-ERK were measured by Western Blot. The mRNA expressions of iNOS, TNF-α, IL-6 and IL-1β were analyzed by RT-qPCR. The results showed that Pseurata H (10, 20, 40 μmol/L) displayed no apparent cytotoxicity in RAW264.7 cells. The release levels of NO, iNOS, TNF-α, and IL-6 were decreased by Pseurata H (10, 20, 40 μmol/L). In addition, Pseurata H also inhibited the expressions of iNOS, COX-2, NF-κB, p-NF-κB, ERK, and p-ERK proteins with concentration effect. Pseurata H has a significant anti-inflammatory effect, and its mechanism of action may be through the inhibition of NF-κB/MAPK signaling pathway to exert anti-inflammatory effects.
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Carboxymethyl pachymaran prevented depressive behaviour induced by sub-chronic stress
E Jiatao, HU Lianhua, WANG Chen, YANG Bin, SUN Dongfang, FANG Zhijia, DENG Qi, SUN Lijun
2024, 41 (2):  44.  doi: 10.3969/j.issn.2095-1736.2024.02.044
Abstract ( 6 )   PDF  
The aim of the study is to investigate the protective effect of carboxymethyl pachymaran on depressive behaviour induced by sub-chronic stress. With the daily exposure to unpredictable mild stress for 2 consecutive weeks, carboxymethyl pachymaran was gavaged in mice at a daily dose of 0.1 g/kg, 0.2 g/kg or 0.4 g/kg for the last 7 days of the modelling procedure. After the experiment, the composition of gut microbiota was investigated by 16S rRNA high-throughput sequencing, and the levels of inflammatory factors in serum were determined by ELISA, the content of short-chain fatty acids in feces was measured by GC-MS, and the depressive behaviour was evaluated by open field test and elevated plus maze test and forced swim test. Compared with the model group, low and middle doses of carboxymethyl pachymaran groups improved depression-like behaviour, and changed the composition of gut microbiota with a decrease in the relative abundance of Gram-negative bacteria (P<0.05). In addition, the middle dose of carboxymethyl pachymaran could lower IL-2 (P<0.05) and IL-6 (P<0.01) levels in serum, and each dose of carboxymethyl pachymaran could down-regulate the levels of isovaleric acid and NE, 5-HT and Ach through modulating Lactobacillus and Parabacteroides relative abundance, thereby reducing the release of IL-6. In conclusion, carboxymethyl pachymaran regulated gut microbiota composition, decreased the release of gut-derived isovaleric acid and NE, 5-HT and Ach, and inhibited depression-associated inflammatory factors IL-6 and IL-2, thereby preventing depression development.
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AITC regulates serotonin secretion and other biological functions in enterochromaffin cells
WANG Siyu, YANG Yali, SI Qiqi, GUO Tailin, HUANG Xinhe
2024, 41 (2):  51.  doi: 10.3969/j.issn.2095-1736.2024.02.051
Abstract ( 7 )   PDF  
In order to investigate the effects of allyl isothiocyanate (AITC) on the synthesis of 5-HT in EC and the biological functions of EC, edible grade AITC were used to intervene with RIN-14B cells (rat pancreatic endocrine cell line), a cell model of EC. The calcium ion concentration in the cells was detected by FLUO-8 AM, the expression level of genes related to 5-HT synthesis was analyzed by qRT-PCR , the levels of 5-HT were determined by UPLC, and the transcriptome of AITC-treated RIN-14B cells was detected and analyzed by RNA-seq and bioinformatics.The results indicated that AITC caused an increase in intracellular calcium ion concentration through activation of TRPA1, while upregulated the expression ofTph1(tryptophan hydroxylase) and Ddc (5-hydroxytryptophan decarboxylase). In addition, GO and KEGG functional enrichment analysis on AITC treated RIN-14B cells showed that AITC mainly regulated glutathione metabolism and relate pathways such as antioxidant and inflammatory regulation, suggesting that AITC may regulate intestinal homeostasis by stimulating EC to promote glutathione metabolism, and also participate in inflammatory regulation of the intestine. The above results provided experimental data and research ideas for further study of the effects of AITC on EC and deeper biological functions of the intestine.
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Isolation, identification and whole genome analysis of a heparin-degrading bacterium
LI Huimin, NING Zhenjing, YUE Siyang, XIE Mingxiu, ZHOU Qihai
2024, 41 (2):  57.  doi: 10.3969/j.issn.2095-1736.2024.02.057
Abstract ( 13 )   PDF  
A bacterium species was isolated from fecal specimens of white-headed langur. This bacterium is non-spore-forming, yellow pigment producing, gram-negative and catalase-positive. It was identified as Sphingobacterium daejeonense NS6-1 based on 16S rRNA gene sequence. NS6-1 can’t hydrolyze starch but is a heparin-degrading bacteria. The heparinase activity of NS6-1 was 46.60 U/mL, and can be increased to 107.29 U/mL in the present of 100 mmol/L Ca2+. Routine drug susceptibility test by K-B disk agar diffusion method showed that NS6-1 was resistant to kanamycin and gentamicin, while sensitive to rifampicin, tetracycline, polymyxin B, ofloxacin, cefoperazone and vancomycin. The genome was 4381042 bp in length with 37.21% overall GC content, including 3852 protein coding sequences. Functional annotation revealed a heparinase Ⅱ/Ⅲ-like protein in CAZy, a aminoglycoside resistance gene in CARD, a gene encoding catechol 1, 2-dioxygenas which can catalyze hydrocarbon-related catabolic in oil. Six gene clusters related to secondary metabolite biosynthesis were predicted by using antiSMASH, of which two were identified as carotenoid synthesis gene clusters and one emulsan synthesis gene clusters. In summary, NS6-1 is a heparin-degrading bacterium, with potential for degradation aromatic hydrocarbons in petroleum and carotenoid synthesis. The present results would lay a theoretical foundation for the further application of the isolated strain.
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Isolation, identification and characterization of a toluene-degrading bacterium
WU Huan, LIN Xiaoying, GUO Tao, NIU Jia, ZHANG Didi
2024, 41 (2):  63.  doi: 10.3969/j.issn.2095-1736.2024.02.063
Abstract ( 6 )   PDF  
To solve the environmental pollution problem caused by toluene emission, a bacterial strain that can use toluene as a single carbon source was screened from the soil near a furniture factory and identified as Pseudomonas sp. M1 by morphological characterization and 16S rDNA analysis. The growth conditions of the strain were optimized by single-factor experiments, and the strain was examined under optimal growth conditions for its ability to utilize different substrates. The growth conditions were optimized by single-factor experiments, and the effects of organic matter utilization and surfactant on the growth and degradation of the strain were investigated under optimal growth conditions. The results showed that strain M1 had strong tolerance to toluene, and the degradation rate reached 63.03% after 3 d at 30 ℃, pH 7.0 and toluene mass concentration of 200 mg/L. In addition to toluene, M1 can also use benzene and ethanol as carbon source and energy source, and can still maintain a certain amount of growth under the combined pollutants of benzene, toluene, ethylbenzene and xylene. The nonionic surfactant TritonX-100 is moderately biodegradable and has the best promotion effect on toluene degradation by strain. At 100 mg/L toluene mass concentration, the toluene degradation rate was 93.35% after the addition of TritonX-100 with 0.5 CMC.
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Knockout of zebrafish kdm4ab affects cold tolerance
ZHANG Mengran, LUO Juntao, ZHANG Yao, HAN Bingshe, ZHANG Junfang
2024, 41 (2):  67.  doi: 10.3969/j.issn.2095-1736.2024.02.067
Abstract ( 20 )   PDF  
o explore the role ofkdm4abin fish under low temperature,kdm4ab-/-zebrafish was constructed by CRISPR/Cas9. RT-qPCR showed that the mRNA expression level ofkdm4abinkdm4ab-/-zebrafish was significantly reduced (P<0.0001), while the expression of kdm4c level was compensatively increased (P<0.05). The detection of zebrafish behavior trajectory tracking system showed that the maximum acceleration ofkdm4ab-/-larval zebrafish was significantly decreased at 18 ℃ (P<0.05), compared with that of WT larval zebrafish. At 8 ℃, the LT50 ofkdm4ab-/-zebrafish was significantly lower than that of WT zebrafish (P<0.0001). After low temperature treatment, the expression of γH2A.X inkdm4ab-/-zebrafish increased significantly, and the level of ROS increased, which indicated that the DNA damage was enhanced. The results showed thatkdm4abhad a certain effect on the cold tolerance of zebrafish, which also provided a new idea for the low temperature tolerance mechanism of fish.
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Genetic diversity analysis of tea insect pest Crenidorsum turpiniae based on mtDNA COI gene partial sequence
MENG Zehong, WANG Weirui, LUO Linli, LI Shuai, PU Yundan, ZHOU Yufeng
2024, 41 (2):  74.  doi: 10.3969/j.issn.2095-1736.2024.02.074
Abstract ( 12 )   PDF  
To investigate the genetic diversity of the tea pest C. turpiniae and ascertain the degree of genetic differentiation and provide a theoretical basis for control, genomic DNA of C. turpiniae populations from 11 different geographical regions in Guizhou Province was extracted. It was amplified through PCR and sequenced in order to determine the mtDNA COI gene fragments. Subsequently, analysis of the gene sequence characteristics, haplotypes and genetic diversity, genetic differentiation and gene flow, molecular variation and haplotype phylogenetic analysis was conducted. The results showed that the mtDNA COI gene sequence fragment amplified from C. turpiniae was 658 bp in length, and the sequence showed obvious A+T bias. A total of 10 haplotypes were defined. The total population of C. turpiniae in Guizhou tea gardens had a high level of genetic diversity, high haplotype diversity (Hd=0.800) and high nucleotide diversity (Pi=0.0670). The total population of the eleven geographical populations had a high degree of genetic differentiation (Fst=0.9203) and low level of gene communication (Nm=0.04). The results of the AMOVA analysis showed the majority of the genetic variation in C. turpiniae population due to differences between groups (FCT=0.9129). The comprehensive analysis of neutral test and mismatch distribution showed that the population had not expanded significantly recently. The results of both haplotype phylogenetic analysis and haplotype network clustering showed that C. turpiniae population was clustered into three clades.
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Scanning electron microscopic observation of sensilla on the antennae of Prionolomia gigas Distant
CHEN Wenhua, YU Shengqing, CAO Kaihong, ZHANG Jing, CHUAN Zhutao, LIU Qing
2024, 41 (2):  80.  doi: 10.3969/j.issn.2095-1736.2024.02.080
Abstract ( 11 )   PDF  
In order to understand the relationship between antennal sensilla of Prionolomia gigas Distant and their olfactory sensory functions, scanning electron microscope was used to examine the types, ultrastructure and distribution patterns of the antennae sensilla from the male and female adult of P. gigas. The results showed that the antennae of P. gigas were filiform and formed of scape, pedicel and flagellum. The length of each segment and the total length of antennae of male and female adults were different. Six sensilla types were observed on antennae of female and male adults, including trichodea sensilla, chaetica sensilla, basiconca sensilla, cavity sensilla, coeloconic sensilla and bent-tipped sensilla. The type of trichodea sensilla was the most numerous and had the widest distribution on antennae. Sensilla in the flagellum were much more abundant than that on scape and pedicel. There was obvious dimorphism that the female of P. gigas had more sensilla types than the male.
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Comprehensive evaluation of the effects of different fertilization measures on the growth of Pueraria lobata in rocky slope covered platform area
QI Li, WU Jilong, LI Xiaolong, HUANG Tao, SUN Qingye
2024, 41 (2):  85.  doi: 10.3969/j.issn.2095-1736.2024.02.085
Abstract ( 3 )   PDF  
In order to obtain a soil matrix improvement measure which is beneficial to the rapid growth of Pueraria lobata in the soil covered platform area of the limestone mine, the rock slope which has been covered with topsoil in the third stage platform area of the east mining area of Yangjialing limestone mine in Wuwei,Anhui, China, was taken as the test area to evaluate the growth of P. lobata on the reconstructed soil under different fertilization measures. Experimental plot settings: A(100% compound fertilizer), B(75% compound fertilizer +25% organic fertilizer), C(50% compound fertilizer +50% organic fertilizer), D(25% compound fertilizer +75% organic fertilizer), E(100% organic fertilizer), CK (control, no fertilizer). In August 2022, physiological and biochemical indexes of P. lobata were analyzed, and the growth status of P. lobata was comprehensively evaluated by entropy weight method. The results showed that compared with CK, fertilization groups increased plant length and leaf dry weight, water content, chlorophyll content, nitrogen, phosphorus and potassium content, soluble protein content and activities of three antioxidant enzymes (POD, CAT and SOD), while decreased MDA content in leaves. The comprehensive evaluation results showed that 25% compound fertilizer +75% organic fertilizer treatment was the most beneficial to the growth of P. lobata, which could be used for the production practice of vegetation reconstruction on the rocky slope.
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Structure and function of chitinase gene in crustaceans
ZHANG Di, JIN Xin, CHENG Runqiang, GENG Changsheng, LI Ye
2024, 41 (2):  91.  doi: 10.3969/j.issn.2095-1736.2024.02.091
Abstract ( 7 )   PDF  
The classification of chitinases in 50 crustaceans was reviewed and it was verified that chitinases in crustaceans could be subdivided into group 1-group 7 according to the classification of amino acid homology. The structure and the function of chitinase in crustaceans were focused in this review. In terms of structure, the characterization of conserved motifs based on amino acid sequence, as well as the composition profile of several domains of group 1-group 7 chitinases were discussed. Then, the three-dimensional structure of one of chitinases from P. trituberculatus, which was considered as a symbol of the crustacean chitinase, was predicted. The functions of crustacean chitinase were further stated and it generally participated in the digestion of chitinous food, in the digestion of old shells during molting, as well as degrading the chitinous cell wall of pathogen to exert its function on immune defense.
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Review of the functions of exosomes in aquatic organisms
CAI Chengjian, JIANG Jiachao, LUO Ping, WANG Lizhang, ZHANG Yuanyuan, GU Xiujun
2024, 41 (2):  97.  doi: 10.3969/j.issn.2095-1736.2024.02.097
Abstract ( 11 )   PDF  
Exosomes are small, membrane-bound vesicles with a diameter ranging from 30 to 150 nm. They are actively released by diverse cell types and exhibit sustained secretion under both physiological and pathological conditions of tissue cells. Exosomes can be found in various body fluids and exert their influence on recipient cells through the delivery of proteins, lipids, nucleic acids, and other bioactive molecules. These molecular cargos enable exosomes to modulate and alter the behavior of target cells, thereby playing significant roles in intercellular communication and immune regulation, among other crucial cellular processes. This article took aquatic-derived exosomes as a focal point, providing a comprehensive synthesis of their compositional characteristics and elucidating the functional diversity of exosomes that observed across different aquatic organisms. Moreover, it undertook an in-depth analysis of the environmental factors impacting the secretion of exosomes, aiming to unravel the intricate relationship between exosome release quantity and these environmental cues. This review presented novel research insights for the study of aquatic-derived exosomes in the context of environmental conservation.
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Establishment and application of a multiple allele-specific PCR method based on a novel high-fidelity Taq polymerase
HEN Zejia, XU Yimeng, SHI Jingjing, ZHOU Yuxun, LI Kai, XIAO Junhua
2024, 41 (2):  103.  doi: 10.3969/j.issn.2095-1736.2024.02.103
Abstract ( 4 )   PDF  
This technology was established on the basis of three selected SNP loci for preliminary testing on mice with partially replaced fragments on chromosome 8, including specificity testing of high-fidelity Taq enzymes and evaluation of the effect of primer concentration on genotyping results as well as sensitivity assessment. It was used to detect nine SNPs in different wild-type chromosome 1 substitution mouse samples. The study showed that the detection results without introducing mutations into primers were consistent with sequencing results. The dilution experiments indicated that the optimal concentration was at least 0.003-0.006 μmol/L, while sensitivity detection revealed that the minimal concentration detection limit could reach below 0.07 ng/μL. Finally, five standard samples’ genotypes representing combinations of nine SNPs were detected. The overall process for applying this highly specific Taq enzyme in multiple allele-specific PCR experiments was eventually validated and established. This article offered a low-cost solution with high specificity and the ability to detect multiple SNP loci, which preserved the reference value for the development of multiple allele-specific PCR technology.
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Optimization of the isolation and culture system of Qinchuan cattle fibroblasts
FENG Xianzhou, CHEN Hui, JIA Yonghong, LIU Songqi, WEN Fei, GUO Songmao, HAN Shuaiqi, HU Jianhong
2024, 41 (2):  110.  doi: 10.3969/j.issn.2095-1736.2024.02.110
Abstract ( 12 )   PDF  
To establish the isolation and culture system of Qinchuan cattle somatic cells, this study used three different methods to extract fibroblasts from the ear margins of Qinchuan cattle. Results showed that fibroblasts could be obtained after 17 d of tissue block culture, while the double enzyme digestion method could be cultured for 9 d for passaging. Compared with that of the enzymatic digestion method Ⅱ(0.9×106/mL), the concentration of fibroblasts obtained by the enzymatic digestion method I(1.24×106/mL)was high and the difference was significant. Third-generation fibroblast immunofluorescenceand flow cytometry identification results showed cell purity of over 95%. The purified fibroblasts were cultured until the logarithmic growth phase at 3 d, and the proliferation rate decreased at 8 d and entered the plateau phase. In conclusion, the dual enzyme digestion method I (0.25% trypsin digestion for 30 min and 1% collagenase for 60 min) was the best method for isolation and culture of Qinchuan cattle ear margin fibroblasts.
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Exploring and practice the construction model of offline first-class courses of farming system #br#
WANG Dongxia, YANG Lina, SONG Xiaoke, XU Yunzhang
2024, 41 (2):  116.  doi: 10.3969/j.issn.2095-1736.2024.02.116
Abstract ( 10 )   PDF  
This paper focused on the development of an offline first-class course teaching model tailored to the core farming science courses at Qinghai University, along with the specific requirements for agricultural and forestry education and talent cultivation. Integrating the current status of the farming system courses in agronomy majors, the demands of offline first-class course construction were combined with practical experiences to analyze the existing issues and deficiencies in course teaching. Taking the necessity and model of constructing offline first-class courses also, the infusion of ideological and political elements was introduced into the curriculum. Additionally, a diversified evaluation model was established for teaching effectiveness. Based on this analysis, the reform strategies and objectives for farming system courses offline first-class course were constructed, then implemented and promoted in practice. It provided guarantee for the construction of first-class agricultural majors, aiming to offer robust support for elevating the overall quality of talent cultivation in Qinghai University.
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The teaching innovation and practice of“comprehensive biology experiment”
XIA Xian, AO Bo, SHI Xiaoshan, FU Shan, GUO Qian, WANG Weidong, TU Junming
2024, 41 (2):  120.  doi: 10.3969/j.issn.2095-1736.2024.02.120
Abstract ( 10 )   PDF  
“Comprehensive biology experiment” is an offline experiment course of our university, the project database covers animals, plants, microorganisms, ecology, toxicology, food and multidisciplinary comprehensive project-based experiments. "Isolation and identification of microbial selenium nanoparticles strains and application of biological selenium nanoparticles" is one of the experimental projects in the project database. This project was based on microbiology (microbial isolation, morphological observation, optimization of fermentation conditions) and molecular biology (DNA extraction, PCR amplification, sequencing analysis), and the knowledge of related disciplines with materials science (biomaterials extraction and morphology analysis) and environmental chemistry (adsorption kinetics analysis), designed as a comprehensive research biology experiment for undergraduate teaching. The experimental project realized the interdisciplinary and integration of courses. It was close to the front line of scientific research and promoted the cultivation of innovative talents. Students designed experimental programs based on the knowledge they have learned and the literatures to promote their independent learning. The project emphasized the process of assessment to mobilize students’ learning enthusiasm. This study took the experimental project as an example and combined with the actual teaching inside and outside the classroom. It explored the construction path of the first-class offline course of “comprehensive biology experiment” to optimize the teaching effect of the course and continuously improve the course.
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Teaching practice and exploration of scientific research program achievements in general education courses
YAN Zhen, YANG Jian, XIONG Li, FAN Jia, ZHANG Dawei, LIN Honghui, LIU Huanhuan
2024, 41 (2):  126.  doi: 10.3969/j.issn.2095-1736.2024.02.126
Abstract ( 18 )   PDF  
The experimental project of "searching for others-application of nucleic acid detection" was summarized here, and the discussion of practical experience of applying part of the research project to the undergraduate general education course was performed. In this experiment project, the Arabidopsis thaliana T-DNA insertion mutants were used as materials to explore the application of nucleic acid detection technology in scientific research and medical testing through genomic DNA extraction, PCR detection and band analysis, covering a complete scientific research process. The practice of this experiment enables non-biology majors to understand the implementation process of biology scientific research projects while acquiring knowledge in biology and experimental skill points related to nucleic acid extraction in biology. It encourages students to think about the background, process and significance of the development of technical means in the nucleic acid testing, cultivating dispersive thinking, innovation and practice ability of students, and laying the foundation for the cultivation of comprehensive and innovative talents in the new era.
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