Table of Content

18 April 2023, Volume 40 Issue 2 Previous Issue    Next Issue

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Discoidin domain receptor 1 (DDR1): a new drug target for solid tumors SHU hang, JIN Tengchuan, WANG Hua 2023, 40 (2):  1.  doi: 10.3969/j.issn.2095-1736.2023.02.001 Abstract ( 320 )   PDF   Save DDR1, a member of the discoidin domain receptor family (DDRs), is widely expressed in human normal tissues and is closely related to the genesis and development of tumors. Based on the structure and biological characteristics of DDR1, this paper reviews the research progress of DDR1 in tumor, expounds the effects of DDR1 on tumor cell proliferation, survival, migration, invasion, epithelial interstitial transformation, and cell metabolism, as well as the important role of DDR1 in tumor microenvironment, and summarizes the progress of anti-tumor drugs targeting DDR1. The study aims to provide theoretical basis and clinical reference for DDR1 as a new target for solid tumor therapy in the future. Related Articles | Metrics

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SREBP1c-ACCα/FAS and SREBP1c-FABP3 axially regulate intracellular lipid synthesis and transport in HepG2 FU Changzhen, ZHENG Ying, LU Yao, WANG Renjun, LIU Qingping 2023, 40 (2):  9.  doi: 10.3969/j.issn.2095-1736.2023.02.009 Abstract ( 375 )   PDF   Save SREBP1c is a key regulator of de novo synthesis and intracellular transport of long-chain fatty acids (LCFAs). In this study, the potential relationship between disruption of the synthesis /transport of LCFAs and non-alcoholic fatty liver disease (NAFLD), which was axially regulated by SREBP1c-ACCα/FAS and SREBP1c-FABPs, was investigated. The adenovirusAd-SREBP1c was prepared for mediating the overexpression of SREBP1c, and the intracellular triglyceride (TG) content was measured by enzymatic assay followed by transfection of HepG2 cells with Ad-SREBP1c. The transcription and translation levels of ACCα, FAS, FABP3 and FABP4 were measured by RT-PCR and Western blot respectively. Ad-SREBP1c virus titer was 1.6×109GFU/mL. The transcription and translation levels of SREBP1 were enhanced by 89.73-fold (P<0.01) and 7.27-fold (P<0.01), respectively, 24 h after transfection in HepG2 cells mediated by Ad-SREBP1. The mRNA levels of ACCα and FAS for downstream LCFAs synthesis were increased by 1.55-fold (P<0.01) and 3.42-fold (P<0.01), respectively, and with increasement each by 1.23-fold (P<0.05) and 1.43-fold (P<0.01) in protein expression levels. The FABP3 mRNA and protein levels were elevated by 4.03-fold (P<0.01) and 2.06-fold (P<0.01) respectively, while no significant change was observed in FABP4. Meanwhile, intracellular TG contents of HepG2 cells of 24 h and 48 h after transfection by Ad-SREBP1c were increased by 1.24-fold (P<0.05) and 2.41-fold (P<0.01), respectively. SREBP1c-ACCα/FAS could axially regulate the de novo synthesis of LCFAs, and SREBP1c-FABP3 could axially mediate the intracellular transport of LCFAs at the same time, which was probably the key mechanism contributing to the development of NAFLD due to cytosolic ectopic lipid deposition. Related Articles | Metrics

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The research of NRF2 in regulating clock gene oscillations under simulated microgravity conditions HUANG Chao, CHEN Hailong, WANG Chunyan, JI Guohua, LYU Ke, YUAN Yanhong, SONG Bo, QU Lina 2023, 40 (2):  14.  doi: 10.3969/j.issn.2095-1736.2023.02.014 Abstract ( 126 )   PDF   Save To investigate whether the simulated microgravity effect affects the expression of CLOCK and BMAL1 through NRF2, the clinostat was used to simulate the microgravity effect. The results showed that the simulated microgravity effect significantly reduced the expression level of NRF2 and significantly increased intracellular ROS level. The expression of CLOCK and BMAL1 decreased significantly, the oscillation rhythm was weakened, the period was shortened, and the phase was changed. Then overexpression of NRF2 could reverse the rhythm changes caused by microgravity effect, and the expression rhythm of CLOCK and BMAL1 was enhanced. The results suggested that the simulated microgravity effect might regulate the expression oscillation of CLOCK and BMAL1 through NRF2. Related Articles | Metrics

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Expression, purification and characterization of human superoxide dismutase 1 and itsmutants YANG Fangyao, JIAN Tiantian, JIANG Lixiang, ZHANG Yu, HUANG Xinhe 2023, 40 (2):  20.  doi: 10.3969/j.issn.2095-1736.2023.02.020 Abstract ( 165 )   PDF   Save SOD is an important functional protein and industrial enzyme, and its source is a hot topic in the field. Here hSOD1 and bosgrunniens SOD1 were analyzed by bioinformatics firstly, and Mt1SOD1 (E25G, P29T, E101V, C112S) and Mt2SOD1 (I18T, N20H, K31V, C112S) were obtained by site-directed mutagenesis.Condons-optimized hSOD1, Mt1SOD1, Mt2SOD1, and bosgrunniens SOD1 were totally synthesized and then cloned into pET-28a(+) to achieve recombinant plasmid for transforming into E. coli BL21(DE3). The four kinds of SOD1 were induced and expressed successfully under the conditions of 1 mmol/LIPTG, 800 μmol/L Cu2+ and 20 μmol/L Zn2+ in LB medium at 25 ℃ and 180 r/min for 16 h. Next, Ni-NTA was used to purify four recombinant SOD1s. The activity of hSOD1 obtained from 100 mL LB was 71094 U/mg, and the yield was 4.57 mg. The activity of Mt1SOD1 was 128506 U/mg, and the yield was 3.13 mg. The activity of Mt2SOD1 was 58700.1 U/mg, and the yield was 5.47 mg. The activity of bos grunniens SOD1 was 42969.5 U/mg, and the yield was 6.81 mg. The enzymatic activity of hSOD1 and Mt1SOD1 remained no change between 25 ℃ and 55 ℃, and the relative enzymatic activity of hSOD1 remained about 50% after incubation at 75 ℃ for 30 minutes, and that of Mt1SOD1 remained 30%. Under the condition of pH 3.6-10.4 for 30 min, both SOD1s can maintain more than 70% of the enzyme activity. hSOD1 and Mt1SOD1 with high activity and high stability were obtained by site-directed mutagenesis, which laid the foundation for the application of SOD1 in medical, health care, cosmetics and other fields. Related Articles | Metrics

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Cloning, expression and analysis of phase transition characteristics of hydrophobic elastin-like polypeptides WANG Shanshan, LIU Yang, GAO Qi, WANG Ling, LU Hongzhao, ZHANG Tao 2023, 40 (2):  27.  doi: 10.3969/j.issn.2095-1736.2023.02.027 Abstract ( 136 )   PDF   Save Elastin-like polypeptides (ELPs) as novel thermal-sensitive biomaterials have a broad application prospect in drug delivery, tissue engineering and protein isolation and purification. It is essential for precise control and rapid biosynthesis of ELPs with specific phase transition characteristics. Technology of recursive directional ligation by plasmid reconstruction was used to successfully construct recombinant plasmids pET28-ELP25, pET28-ELP50 and pET28-ELP100 as valine as guest amino acid of ELPs. Then, the recombinant plasmids were transformed into Escherichia coli BL21(DE3), respectively. After cultures of recombinant strains under self-induced medium, SDS-PAGE results showed that ELPs were successfully soluble over-expressed in E. coli system, and the expression level was over 30% of total cell proteins. ELP25, ELP50 and ELP100 were separated and purified by three rounds of inverse transition cycling (ITC). Thermal turbidimetry was used for measurement of phase transition temperature (Tt) of ELPs. Analysis of phase transition characteristics revealed that the Ttof ELPs was negatively correlated with polypeptide concentration, ionic strength and molecular weight of ELPs. The results provide a theoretical basis for further recombinant expression of thermal-sensitive polypeptide biomaterials with required phase transition characteristic. Related Articles | Metrics

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Decolorization of several different dyes by Trametes sp. 4281 laccase LI Li, ZHANG Xuecheng, XIAO Yazhong 2023, 40 (2):  33.  doi: 10.3969/j.issn.2095-1736.2023.02.033 Abstract ( 135 )   PDF   Save Trametes sp. 4281 is a new type of highly efficient laccase production strain independently screened and identified in the laboratory.In order to explore the potential of laccase to decolorize dye, crude Trametes sp. 4281 laccase solution was used to decolorize three categories of dyes, i.e. azo, anthraquinone and phenylmethane. The effects of different mediums, temperatures, pHs, medium concentration, enzyme amount, reaction times and metal ions on the dye decolorization were investigated. The results showed that addition of mediators had a significant influence on the decolorization efficiency; the six dyes were almost completely decolorized within 8 h, with the decolorization rates of reactive blue, alizarin red, methyl orange and crystal violet under the optimum reaction conditions reaching 93.9%, 92.3%, 90% and 90%, and those of Congo red, fuchsin reaching 76.6% and 75%, respectively. Mn2+ and high concentrations of Fe2+ had inhibitory effect on the decolorization, while in the presence of metal ions of Na+, Mg2+, Cu2+ Ni2+, and Zn2+ the laccase remained high decolorization rates to the dyes. The results suggest that Trametes sp. 4281 laccase has great application potential in the dye decolorization, and has a good prospect in the treatment of environmental wastewater. Related Articles | Metrics

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Bacteria diversity in Daihai Lake in Inner Mongolia JIANG Kai, CAO Chunling 2023, 40 (2):  38.  doi: 10.3969/j.issn.2095-1736.2023.02.038 Abstract ( 148 )   PDF   Save Bacterial diversity in water and sediments of Daihai Lake was analyzed by culture method and metagenomic sequencing in this study. The bacteria isolated by culture method belonged to 4 phyla, 6 classes, 18 orders, 25 families, 47 genera, 77 species, including 20 potential novel species, the dominant phyla were Proteobacteria and Firmicutes. It was found that 0.22 μm filtration can remove most common bacteria and improve the detection of novel species. A total of 83 phyla were identified by metagenomic sequencing. Proteobacteria and Bacteroidetes were the dominant phyla. Metagenomic sequencing identified one unclassified phyla and 50 candidate phyla, of which 33 were candidate phyla radiation (CPR). The results showed that there are high abundance and diversity of bacteria, and abundant CPR strains in Daihai Lake. The results laid a foundation for further research on microbial resources mining and ecological functions in Daihai Lake. Related Articles | Metrics

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Distribution of Tuber in Liupan mountain and its ecological significance DENG Xiaojuan, LIU Peigui, CHEN Juan, QIAO Peng, LIU Jianli, AMAT Rustam, YAN Xingfu 2023, 40 (2):  45.  doi: 10.3969/j.issn.2095-1736.2023.02.045 Abstract ( 106 )   PDF   Save The resources of Tuber were investigated in Liupan mountain in 2019 and 2020. Twelve specimens of Tuber were collected and identified. After morphological observation and molecular phylogenetic analysis, the specimens were identified as T. lishanense, T. pseudohimalayense and T. umbilicatum, in which T. pseudohimalayense is with great economic value. Based on collected specimens, the morphological characters and variation of these three species were described and discussed in details. The results broaden distribution area of Tuber in China. The record and ecological data of Tuber in Liupan mountain were supplemented. It is the first time to confirm the distribution of these three Tuber spp.in Liupan mountain. The results are significant to the resources diversity, biogeographic and ecological study of Tuber in China. Related Articles | Metrics

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Composition characteristics of aerial algae and their influencing factors in different forests in Spring WEI Liman, SUN Qingye, ZHAO Qiong, QI Li 2023, 40 (2):  51.  doi: 10.3969/j.issn.2095-1736.2023.02.051 Abstract ( 116 )   PDF   Save The species composition of soil algae and corticolous algae of Liquidambar formosana, Cyclobalanopsis glauca, Pinus massoniana and Cunninghamia lanceolata in Ningguo city, Anhui Province was conducted in March 2021, the richness of aerial algae and its relationships with environmental factors were analyzed. The results showed that a total of 83 species of algae were detected in the surface soil and bark, belonging to 5 phyla, 7 classes, 21 orders, 35 families, 45 genera, dominated by Cyanophyta，Chlorophyta respectively. The richness of aerial algae in different attachment substrates was soil >bark, 22 species were present on both substrates. The dominant families of soil algae and corticolous algae were Ulotrichaceae of Chlorophyta and Chroococcaceae of Cyanophyta, respectively. The richness of soil algae was as follows, Cyclobalanopsis glauca > Liquidambar formosana > Cunninghamia lanceolata > Pinus massoniana, and the most significant correlation factors affecting its richness were ammonia nitrogen and organic matter (P<0.05). The richness of corticolous algae was as follows, Pinus massoniana > Cyclobalanopsis glauca > Cunninghamia lanceolata > Liquidambar formosana, and was not significantly affected by surface bark nutrients. Related Articles | Metrics

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Effects of LED mixed light on growth and phycocyanin accumulation of Spirulina XIE Jiawen, XU Peilun, HAN Pei, QIAN Jun, ZHANG Peidong, WANG Bang, GUO Lingyue, ZHOU Wenguang 2023, 40 (2):  58.  doi: 10.3969/j.issn.2095-1736.2023.02.058 Abstract ( 178 )   PDF   Save To explore the effects of different light equality on growth and phycocyanin accumulation of Spirulina, the algal cells were cultured in three monochromatic lights (i.e.white (W), blue (B), red (R) light) and their nine mixed light (mixed light intensity ratios) using light emitting diode (LED) as light source. Simultaneously, the algal yield, phycocyanin content and productivity were measured during the experimental period. The results showed that the red group promoted the growth rate of Spirulina, while the blue group contributed to improve both of phycocyanin content and purity in algal cells. Algal yield as optical density (OD) and dry weight in B+R (75%+25%) group increased by 154% and 157% compared with that in the blue group, whereas phycocyanin content and purity increased by 43% and 22% compared with that in the red group, respectively. OD value and dry weight in B+W (75%+25%) group were improved by 107% and 119%, respectively, compared to that in the blue group, but each of phycocyanin content and purity was almost same as that in blue group. The phycocyanin productivity in W+R (50%+50%) group was highest, 42 mg/（L·d）, among the tested groups. Related Articles | Metrics

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Correlation analysis and QTLs mapping of lodging resistance and yield traits in rice HAN Leifeng, ZHOU Ran, ZHOU Tao, LIN Cuixiang, GAN Quan, NI Dahu, SHI Yingyao, SONG Fengshun 2023, 40 (2):  65.  doi: 10.3969/j.issn.2095-1736.2023.02.065 Abstract ( 118 )   PDF   Save With high-yield varieties Yangdao 6 and Lvhan 1 build 192 recombinant inbred lines as the experimental material, the eight lodging resistance traits (plant height, fresh weight, bending force, base transverse area, base area of the medullary cavity, basal stem wall area, wall thickness and lodging index) and three yield related traits (spike length, weight per panicle and effective panicle number) were selected for correlation analysis and QTL mapping. A total of 14 antiinversion related QTLS were identified, which were distributed on chromosome 1, 2, 3, 5, 7, 8, 10 and 12. LOD ranged from 3.02 to 14.78, additive effect ranged from -5.22 to 2.36, and QTL contribution rate was 3.36% to 20%. Eight yield related QTLS were distributed on chromosome 1, 3, 5 and 12, with LOD ranging from 3.1 to 9.43, additive effect value ranging from -12.55 to 4.58, and QTL contribution rate ranging from 3.93% to 14.53%. There was significant negative correlation between yield traits and lodging resistance, and many QTLs appeared in clusters. In this study, QTL mapping and correlation analysis were conducted for lodging resistance traits and yield traits, and the projection area method was adopted instead of wall thickness as an evaluation index to comprehensively excavate excellent QTL loci, providing scientific basis for breeding high yield and lodging resistance rice varieties. Related Articles | Metrics

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Effects of zinc on growth and physiological indices of potato plantlets under drought stress #br# DANG Weiying, TANG Xun, ZHAO Jing, ZHAO Guibin, ZHU Yongyong, ZHANG Ning, SI Huaijun 2023, 40 (2):  71.  doi: 10.3969/j.issn.2095-1736.2023.02.071 Abstract ( 143 )   PDF   Save In order to elucidate the effect of zinc on the growth and development of potato under drought stress, the tissue culture seedlings of potato cultivar “Atlantic” , and the medium with different zinc contents (0, 8.6, 17.2, 25.8, 34.4 and 43 mg/L) were used as the test material. After cultured for 15 d, the medium was replaced with medium containing 20% PEG6000 to simulate drought stress for 15 d. The phenotype, antioxidant enzyme activity and osmotic adjustment substance content of the tissue culture seedlings were determined, and the drought resistance was analyzed. The results showed that zinc had a significant effect on the growth of potato tissue culture seedlings under PEG stress. The tissue culture seedlings performed best when the zinc content of the medium was 17.2 mg/L. Among them, the fresh weight and plant height increased by 50% and 34.66% respectively when the zinc content of the medium was 17.2 mg/L compared with that of the normal MS treatment; the activities of catalase (CAT), superoxide dismutase (SOD) and Pro Compared with that of the normal MS treatment, the content increased by 9.88%, 10% and 3.88% respectively, and the MDA content decreased by 21.86% compared with that of the normal MS treatment. The results showed that exogenous application of an appropriate amount of zinc could promote the growth and development of tissue culture seedlings, and when the zinc content of the medium was 17.2 mg/L, the drought tolerance of potato tissue culture seedlings could be significantly improved. The results provided a basis for further elucidating the mechanism of zinc regulation of potato drought resistance. Related Articles | Metrics

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Fe(II)-oxidizing bacterium promotes the formation of iron plaque and the prevention of Cd from soils to rice by iron plaque CHEN Jinyuan, JIN Mengnan, FAN Fangjuan, MA Shaochen, LI Sujun, SUN Riyang, LI Ding 2023, 40 (2):  75.  doi: 10.3969/j.issn.2095-1736.2023.02.075 Abstract ( 148 )   PDF   Save Through the ferrous sulfide gradient tube and phenanthroline spectrophotometric experiments, the ability of Fe(II)oxidation for strain LLDRA6 was explored. By using methods such as scanning electron microscope-energy spectrum analysis (SEM-EDS), dithionite-citrate-bicarbonate (DCB) extraction test, and the measurement of Cd content in two rice varieties, the effect of strain LLDRA6 on the formation of IP and its effect on the migration of Cd from soils to ricewere discussed. The results showed that strain LLDRA6 formed an obvious Fe(II) oxidation circle in the ferrous sulfide gradient tube. In the presence of strain LLDRA6, the Fe(II) oxidation rate of LB liquid medium was higher than that of LB liquid medium without LLDRA6 (the control group), suggesting that this strain was an Fe(II)-oxidizing bacterium(FeOB); SEM-EDS results showed that, after addition of strain LLDRA6 and Fe(II) ions in soils, layered precipitates were formed on the surface of the rice roots, and the energy spectrum analysis (EDS) demonstrated that a large amount of Fe element were contained in these precipitates; the DCB extraction test showed that after treatment with strain LLDRA6 and Fe(II), the Fe content of IP was significantly higher than that of IP formed in other treatment groups, indicating that the strain facilitated the formation of IP. Moreover, the Cd content of the stems, leaves and grains were significantly lower than that of corresponding parts of rice grown in other treatment groups. These results demonstrated that strain LLDRA6 benefited to form IP around rice root surfaces, and the IP had a significant blocking effect on the migration of Cd from soils to rice. Related Articles | Metrics

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Characteristics of the gonadal transcriptiome of intersex red claw crayfish Cherax quadricarinatus LI Fei, ZHENG Jianbo, CHENG Shun, LIU Shili, LIU Yinuo, JIA Yongyi, CHI Meili, GU Zhimin 2023, 40 (2):  80.  doi: 10.3969/j.issn.2095-1736.2023.02.080 Abstract ( 106 )   PDF   Save In order to study the difference between the intersex and normal individuals, their testis and ovary were sequenced with Illumina Novaseq 6000 in this study, and were analyzed and compared in biological information and gene expression level. With sequencing, 83.79 Gb clean reads were obtained in total, and 6.29 Gb on average, the Q30 bases percentage was more than 94.25%. After assembling, 53077 Unigenes and 73588 transcripts were obtained, then they were mapped with the six international data, the mapping ratio was 18.17%-36.07%. The gene expression analyzing indicated that there were 3543 Unigenes co-expressed, the correlation of expression between ovaries was the greatest, the correlation for testis was followed, the correlation between ovary and testis was the last one. There were differences in the amount of expression gene between normal and intersex individuals, with 310 genes in total. In conclusion, besides differences in morphology between intersex and normal individuals, there were differences in transcription level for gonads, and the level was different for different kinds of intersex individuals. Related Articles | Metrics

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Comparison on genes expression of AQP1 and AQP3 and activity of digestive enzymes in small intestine between hibernating and non-hibernating Daurian ground squirrels ZHONG Qiumei, WANG Jianli 2023, 40 (2):  86.  doi: 10.3969/j.issn.2095-1736.2023.02.086 Abstract ( 126 )   PDF   Save The Daurian ground squirrel (Spermophilus dauricus) is a lipid storage hibernating animal, as a result, it does not eat any food and drink water during hibernation. However, it is not well known what the differences are in genes expression of Aquaporin (AQP) and activity of digestive enzymes between hibernating and non-hibernating periods. To investigate the adaptive response of small intestine at physiological and biochemical levels in hibernating rodents, the genes expression ofAQP1andAQP3and the activity of digestive enzymes (protease, sucrase, maltase and lipase) in the ileum of Daurian ground squirrels were examined during active (summer, July), hibernating (winter, December) and arousal (spring, March) periods. The results indicated the expression level ofAQP1gene in hibernating period was higher than that in active and arousal periods, while the expression level ofAQP3gene in hibernating period was lower than that in active and arousal periods. Total protease activity was higher in active period than that in hibernating and arousal periods. The activity of maltase and sucrase in hibernating period was lower than that in arousal period. The activity of lipase was relatively stable in these three periods.These results indicate that the ground squirrel regulates the water balance and nutrient by changing the AQP genes expression and the some enzymes activity of small intestine, which could acclimate to its hibernation habit and variations in energy demands. Related Articles | Metrics

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Research progress on mechanism of African swine fever virus regulating host immune response YUAN Li, XIONG Dan, KANG Xilong, PAN Zhiming, JIAO Xin’an 2023, 40 (2):  91.  doi: 10.3969/j.issn.2095-1736.2023.02.091 Abstract ( 163 )   PDF   Save African swine fever virus (ASFV) encodes a variety of immune escape proteins to antagonize the host immune response, resulting in the persistent infection of the virus in the host. The main difficulty in the development of African swine fever (ASF) vaccine is that little is known about the virulence related genes, pathogenicity and immune regulation mechanism of ASFV. This paper summarized the immune regulation mechanism of African swine fever virus (ASFV) after invading the host. It discussed the specific function of ASFV immune-regulation proteins from the perspective of innate immunity and acquired immunity: regulating inflammation, interferon production, antigen presentation and cellular immunity, and explored potential new vaccine candidates of ASFV: recombinant NH/P68 attenuated strain, new fusion protein ZPMT, gene deletion strain ASFV-ΔECM3, etc, so as to provide reference for the study of ASFV pathogenesis and the development of new vaccines. Related Articles | Metrics

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Research progress of UDP-sugar biosynthesis CHEN Guihang, LI Chun, FENG Xudong 2023, 40 (2):  95.  doi: 10.3969/j.issn.2095-1736.2023.02.095 Abstract ( 993 )   PDF   Save Uridine diphosphate(UDP)-sugar is an important type of sugar donors for glycosylation modification. The in vivo synthesis of UDP-sugar was summarized from three aspects: synthase pathway, phosphorylase pathway and kinase pathway. Due to the lack of key enzymes, only UDP-glucose (UDP-Glc) could be quickly obtained from the above three pathways. At the same time, with UDP-glucose as the starting material and UDP-glucuronic acid (UDP GlcA) as the intermediate, the rapid interconversions between UDP-sugars could be realized by specific functional enzymes. The latest progress in UDP-sugar synthesis catalyzed by specific functional enzymes was reviewed, and the important roles of dehydrogenase, decarboxylase, isomerase and reductase in UDP-sugar interconversions were discussed. The existing problems of UDP-sugar synthesis were analyzed and the future research direction of UDP-sugar synthesis was prospected, aiming at providing new ideas for tapping the potential of UDP-sugar donors and realizing glycosylation modification with efficiency and low cost. Related Articles | Metrics

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Research progress on NAD+metabolism and its effect on aging-related diseases YANG Honghui, SUN Yuan, WANG Xinbo, YAN Wei 2023, 40 (2):  101.  doi: 10.3969/j.issn.2095-1736.2023.02.101 Abstract ( 212 )   PDF   Save In this work, we reviewed the research progress of NAD+metabolism mechanism and its effect on aging-related diseases, focusing on the signaling pathways and cellular processes involved in NAD+, as well as its role in health and diseases from the aspects of NAD+biosynthesis pathway, consumption pathway and its regulatory mechanism. Based on the mechanism of action of precursors and key enzymes in the NAD+metabolic network, the potential therapeutic approaches of NAD+metabolism regulation on aging-related diseases were discussed, providing new perspectives for understanding the complex causal relationship between NAD+levels decline and aging-related diseases. The issues to be solved in the process of NAD+research and development were summarized from the aspects of the distribution and transportation of NAD+and its precursors, the production and repair of toxic metabolites, and the monitoring and measurement of NAD+metabolic dynamics, and finally, the broad prospects for disease treatment and drug development with NAD+as the target were prospective. Related Articles | Metrics

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Optimization of enzymatic hydrolysis on donkey blood to prepare antioxidant peptides WANG Shuang, LIU Xuesong, JIANG Botao 2023, 40 (2):  109.  doi: 10.3969/j.issn.2095-1736.2023.02.109 Abstract ( 123 )   PDF   Save In this study, the antioxidant capacity of different donkey blood hydrolyzate was compared with DPPH radical scavenging rate and total antioxidant capacity as evaluation index. Through the test results, the hydrolysate with the strongest antioxidant activity was obtained. The enzymatic hydrolysis time, temperature, pH and enzyme-substrate ratio were optimized by single factor test and response surface method, and the actual antioxidant activity was verified by experiment. The final optimized hydrolyzate was ultrafiltered, and the antioxidant activities of different ultrafiltration components were tested. On the basis of single factor test and response surface method, the optimum enzymatic hydrolysis conditions of papin hydrolyzate were 55 ℃, pH 7, time 3.54 h and enzyme substrate ratio 20%. Under this condition, the DPPH radical scavenging rate of the hydrolysate was 71.50% and the total antioxidant capacity was 1.53 mmol/g. After verification, the DPPH radical scavenging rate of the actual enzymatic hydrolysis product was 70.40%±1.33%, and the total antioxidant capacity was （1.51±0.25）mmol/g. The antioxidant acitivity of the ultrafiltration component 3 （<3 ku） was determined to be the strongest, with DPPH radical scavenging rate of 71.26%±5.63% and the total antioxidant capacity of （1.49±0.36） mmol/g. The above results indicated that the donkey blood hydrolyzate treated with papain and its ultrafiltration component had excellent effects on antioxidant activity and radical scavenging. Related Articles | Metrics

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Speculation and practice of the online plus offline blending teaching in cell biology LI Junlin, CUI Jihong, WANG Cuiling, HUANG Xuan, LI Xiang, GUAN Feng 2023, 40 (2):  115.  doi: 10.3969/j.issn.2095-1736.2023.02.115 Abstract ( 176 )   PDF   Save In recent years, in the online plus offline blending teaching of cell biology, it was found that negative performances of resistance and/or improper use, which might affect teaching quality. Combined the traditional teaching paradigm, the achievements of predecessors and our teaching practice, this paper takes the teaching content of "cell death" as an example to summarize the ways to improve the quality of online and offline blending teaching. Firstly, the concept of blending teaching shall be rationally understood and accepted. Secondly, guided under the traditional teaching paradigm, learning and teaching performance shall be analyzed, the training plans shall be formulated, and teaching content shall be reasonably reconstructed. Thirdly, the teaching content shall be refined, and the teaching mode shall be designed and optimized. And finally, sustainable development of the process and multiple assessment system can effectively promote a student-teacher interaction, inspire learning interest, and cultivate entrepreneurial and innovative mindset and abilities. Related Articles | Metrics

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Annexin V-FITC/PI detection of apoptosis experiment WEI Yuan, WANG Honggang, CHEN Chengbin, WANG Chunguo, SONG Wenqin, LI Dengwen, ZHAO Liqing 2023, 40 (2):  119.  doi: 10.3969/j.issn.2095-1736.2023.02.119 Abstract ( 375 )   PDF   Save An experimental course was set up to show how to detect apoptosis with Annexin V-FITC/PI to undergraduate students. Firstly, Etoposide (VP-16) was added to the Jurkat cells to induce the apoptosis, and then the apoptosis was detected by Annexin V-FITC/PI Apoptosis Detection Kit and photographed by fluorescence microscopy. The results showed that viable apoptotic cells showed green fluorescence under UV excitation, while late apoptotic and non-viable non-apoptoic cell showed red fluorescence. The number of cells was normal which had been counted in the control group. The apoptosis rate of different stages of cells increased to drug concentration increased after treated with drugs. The viable apoptotic cells and late apoptotic cells accounted for the highest proportion. While the cells were killed substantially, few early apoptotic cells remained when treated with high drug concentration. This experiment combined the Annexin V-FITC/PI kit with fluorescence microscopy, breaking the limitations of experimental conditions such as teaching funds, scale and instruments. Because this method costs relatively low, it is very suitable for large classes. On the other hand, the results are intuitive, so it is convenient for the students to understand the apoptosis principles and master the assay protocols. It meets the teaching requirements for undergraduate students teaching work, featured by high comprehensive, well-designed and researched. So that this apoptosis assay method effectively improves the level and quality of cell biology experimental teaching work. Related Articles | Metrics

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The creation and practice of the “five fusion” hybrid teaching system of comprehensive experiment of fermentation technology LI Youran, SHI Guiyang 2023, 40 (2):  123.  doi: 10.3969/j.issn.2095-1736.2023.02.123 Abstract ( 124 )   PDF   Save The comprehensive experiment of fermentation technology is the core course for cultivating students’ innovative practice ability in the undergraduate major of biotechnology. With the progress of the discipline and the development of the industry, the original teaching system has problems such as limited content setting, lack of platform support, rigid organizational model, single assessment standard and imperfect experimental system, which affect the cultivation of engineering practice and innovation ability of professional talents. In response to solve those problems, the research and practice of the online and offline hybrid teaching system of the “five-integration” fermentation process comprehensive experiment of bioengineering undergraduates has been carried out, and good results have been achieved. Related Articles | Metrics

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Optimization of the methods for visual determination of viral titer and efficacy of antiviral drugs TANG Qi, CAO Yuanyuan, ZHANG Xinyu, PENG Wangping, LU Baojing, HUANG Shenghai 2023, 40 (2):  127.  doi: 10.3969/j.issn.2095-1736.2023.02.127 Abstract ( 224 )   PDF   Save Experimental course teaching is an important measure to cultivate students’ innovative abilities. In the cell culture course of biomedical majors, the comprehensive experiment of determining viral titer and antiviral drug efficacy is important content. However, there are many problems in the course of traditional plaque experiments. In this paper, the experimental efficiency was first improved by optimizing the traditional steps of plaque assays. By integrating plaque assay and drug assay, the experimental methods of determining the effect of antiviral drugs were simplified. And the experimental skills and scientific interest of students were enhanced through the quantitative and visual determination of viral titers and antiviral efficacy. In addition, the traditional plaque assays neither detect titers of the viruses without significant cytopathic effects (CPE), nor test the activity of antiviral agents to these viruses. In this paper, to address these problems, a cutting-edge technology-fluorescent enzyme-linked immune spots (Fluo-ELISPOT) was applied to visualize and quantify viral plaques and to determine viral titers and efficacy of antiviral drugs, which provided new experimental techniques and new insights for the research and development of antiviral drugs. It also made a positive attempt to optimize the experimental teaching system of medical biology and improve the quality of cell experimental teaching, and further promoted the reform and innovation of student experimental teaching. Related Articles | Metrics