Table of Content

18 June 2020, Volume 37 Issue 3 Previous Issue    Next Issue

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The study of GPI-anchored proteomics in lung cancer cell exosomes WU Wei-bo, LIAO Hai, ZHOU Da-peng 2020, 37 (3):  1.  doi: 10.3969/j.issn.2095-1736.2020.03.001 Abstract ( 447 )   PDF   Save GPI(Glycosylphosphatidylinositol)-anchored proteins are complexes which consist of proteins, lipids and sugar chains. Studies indicate that GPI-anchored proteins are involved in various physiological processes such as immune recognition, cell communication and signal transduction. Exosomes regulate many pathophysiological processes including immune response and tumor growth, which make it significant to research the GPI-anchored proteins over the exosomes. In order to find the GPI-anchored proteins which can be used as the marker of the detection of lung cancer, 114 GPI-anchored proteins, 43 of which are significantly different between lung cancer patients and healthy people were studied. And the data were screened out using the mRNA expression array data of lung cancer patients and healthy people in TCGA (The Cancer Genome Atlas) database. DIA (Data Independent Acquisition) quantitative and Semi-quantitative technology were used to analysis of proteins from exosomes of NCI-H838 non-small cell lung cancer cell line, with total cellular membrane proteins as control, and 15 GPI-anchored proteins from exosomes were screened out. At last, 7 GPI-anchored proteins, including MSLN (mesothelin), CPM (carboxypeptidase M), SMPDL3B(sphingomyelin phosphodiesterase, acid-like 3B), HYAL2 (hyaluronoglucosaminidase 2), EFNA5(ephrin-A5), RECK(reversion-inducing-cysteine-rich protein with kazal motifs) and GPC1(glypican 1), were searched out by the data of GPI-anchored proteins in lung cancer patients and NCI-H838 cell exosomes, which can be used as the markers in the detection of lung cancer. Related Articles | Metrics

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The interactions of MAPK/HOG signal pathway factors in Pichia pastoris WANG Rong-bin, ZHAO Tian-yu, ZHUO Jun-lin, LIU Chun-li, LIU Xiu-xia, YANG Yan-kun, BAI Zhong-hu 2020, 37 (3):  7.  doi: 10.3969/j.issn.2095-1736.2020.03.007 Abstract ( 398 )   PDF   Save MAPK/HOG signal pathway of yeast plays an important role in adapting the change of hyperosmotic stress outside. The current studies on this pathway, however, are practically based on Saccharomyces cerevisiae instead of Pichia pastoris, which limits the application of this excellent heterologous protein expression system. In this study, CRISPR/Cas9 system was applied to knock out the key transcription factor genes PBS2, HOG1, HOT1 and HOT2 in P. pastoris. Then, these mutant strains and GS115 strain were treated both in pre-stress and hyperosmotic condition. The results revealed that HOG1, HOT1 and HOT2 played important roles in protecting cells from hyperosmotic stress. The transcriptional levels of PBS2 and HOG1 were constitutive and they up-regulated MSN2 and HOT1, but no significant changes were found in the transcription level of HOT2. However, MSN2 and HOT1 were still induced by hyperosmotic stress indicating that more transcription factors were involved in the regulation network. Several genes in the MAPK/HOG pathway, including GT1, DOG2, DAK1, HXT1, HSP12 and CTT1, were confirmed. Hog1 was a key transcription factor for up-regulating HSP12 and CTT1. The expression of HXT1 was regulated by Hot2. However, Hot1 was not involved in the regulation network. Finally, a novel MAPK/HOG pathway model in P. pastoris was established. Related Articles | Metrics

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Analysis of gene fusion characteristics based on genome-wide level LI Ying-xia, ZHANG Ting-ting, MA Lei 2020, 37 (3):  12.  doi: 10.3969/j.issn.2095-1736.2020.03.012 Abstract ( 458 )   PDF   Save Gene fusion is one of the important causes of tumorigenesis, and almost half of all malignant tumors have the involvement of fusion genes. Exploring the characteristics of fusion genes has become a hot topic in life science and medical research. In this study, 11 632 gene fusion data were collected from ChiTaRS-3.1 database, from gene fusion, fusion gene distribution across the chromosome, and genes involved in fusion and the characteristics, as well as the analysis of the characteristics of the fusion gene using leukemia as an example. The results showed that gene fusion was dominated by chromosomal translocations, with more frequent mutations on chromosomes 11, 17 and 19 involving a total of 8733 genes, of which only 9% of genes participate in multiple fusion events, and 13% of gene fusion events were associated with leukemia. The distribution of gene fusion in the genome was non-random and concentrated. This study would provide new clues for the subsequent discovery of the mechanism of chromosomal gene fusion and its treatment and prevention of diseases. Related Articles | Metrics

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Construction of an enhanced expression vector and expression of bovine interferon alpha using Corynebacterium glutamicum SUN Man-man, GAO Xiong , FANG Qiu-wu, YAN Hao, LIU Xiu-xia, YANG Yan-kun, BAI Zhong-hu 2020, 37 (3):  16.  doi: 10.3969/j.issn.2095-1736.2020.03.016 Abstract ( 301 )   PDF   Save Corynebacterium glutamicum is a traditional food-grade industrial microorganism which has been developed as a novel endotoxin-free recombinant protein expression host in recent years. To improve the expressing ability of recombinant protein, an endogenous bicistronic element was introduced in front of the multiple cloning site of pXMJ19, bringing the plasmid an enhanced Ptac promoter. The new expression vector was named pSM19 and its expression strength was 99.5% higher than that of pXMJ19 measured by the enhanced green fluorescent protein reporter. The codon-optimized, histidine-tagged BoIFN-α encoding gene was then cloned into the pSM19 and the vector pSM19-BoIFN-α was transformed into the expression strain Corynebacterium glutamicum CGMCC1.15647. When cultured at 30 ℃, the recombinant BoIFN-α protein mostly existed in the form of inclusion bodies, while the solubility of which was greatly improved when cultured at 16 ℃. Scale-up culture was carried out in a 5 L bioreactor in order to obtain more cell pellets containing recombinant protein, followed by a simple purification procedure utilizing the nickel column. The yield of recombinant BoIFN-α was estimated at 68 mg/L and its antiviral activity was (1.35±0.23)×106 U/mg determined by the MDBK-VSV system. Related Articles | Metrics

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Genetic variation and evolution of Pik-2, a resistance gene against rice blast NIU Yu-fang, XIAO Gui, WU Jun, BAI Bin, ZHOU Bo, XIAO Ying-hui 2020, 37 (3):  21.  doi: 0.3969/j.issn.2095-1736.2020.03.021 Abstract ( 149 )   PDF   Save The Pik locus, located on the long arm of chromosome 11, is a hotspot region for rice blast resistance. To study the diversity and distribution of the resistance (R) genes at the Pik locus will be useful for revealing the evolutionary mechanisms of this gene and to further explore its alleles. We sequenced and analyzed the Pik-2 coding region of 400 Oryza glaberrima and 400 Oryza sativa cultivars from different countries and regions. 113 and 78 polymorphic sites were found in O. glaberrima and O. sativa, respectively, which showed high mutation frequency at the nucleotide position 1879. According to the characteristics of the base variations, 17 haplotypes of O. glaberrima and 12 haplotypes of O. sativa were identified. The results indicated that the Pik-2 haplotype distribution in O. glaberrima was characterized by large aggregation and small dispersion, in contrast to the very extensive distribution in O. sativa. Phylogenetic analysis of different haplotypes showed that most groups contained both African and Asian rice haplotypes, suggesting that African and Asian rice had similar haplotypes in the Pik-2 LRR domain during their independent evolution. Related Articles | Metrics

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Analysis of the codon usage preference of genes in midgut of Pieris rapae ZHANG Xian, GUO Shi-qi, WANG Wei-wei, LIAO Hai, ZHOU Jia-Yu 2020, 37 (3):  26.  doi: 10.3969/j.issn.2095-1736.2020.03.026 Abstract ( 274 )   PDF   Save In order to understand the preference characteristics of codon usage in the midgut expression genes of Pieris rapae, and to provide a theoretical basis for application of genetic engineering technology to achieve the heterologous expression of serine protease from P. rapae, in this paper, 51 457 full-length sequences in the midgut transcriptome of P. rapae were calculated and counted for GC content, effective number of codons(ENC), relative synonymous codon usage(RSCU), codon usage frequency and other parameters to measure the codon usage preference of P. rapae by using bioinformatics softwares including CodonW, CHIP and CUSP. The results showed that the total GC content was 40.43%, and the average GC content of the third nucleotide of the codon (GC3) was 38.16%. The distribution of ENC ranged from 22.81 to 61.00 with an average ENC of 53.12. There were 1160 unigenes with ENC less than 35 which accounted for 2.25% of the total. The overall codon preference of genes was not high, but there were still differences in codon usage preference among different genes. The transcriptome codon has obvious preference for codons ending in A or U. The four codons of GAA, UUU, AAU and UAU were the optimal codons for the midgut expression genes of P. rapae. The codon usage preferences of serine proteases (chymotrypsin and trypsin) involved in digestion in the midgut of P. rapae and six different heterologous expression hosts were compared and analyzed. The results showed that Escherichia coli B was the most difficult host to heterologous expression, followed by insects High 5 cells and mammalian HEK293S cells. The expression difficulty of tobacco, Pichia pastoris and Arabidopsis thaliana was relatively low. Related Articles | Metrics

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Constructing and activity verifying of chicken bmp4 overexpression and knockout vectors LI Ting-ting, SUN Chang-hua, ZHOU Shu-jian, JIN Jing, ZHANG Ya-ni, ZUO Qi-sheng 2020, 37 (3):  31.  doi: 10.3969/j.issn.2095-1736.2020.03.031 Abstract ( 306 )   PDF   Save Bmp4 regulates the formation of primordial germ cells (PGCs) through BMP signaling pathway in mammals, but its function and mechanism in the formation of chicken PGCs is still unclear. In this study, overexpression and knockout vectors of chicken bmp4 were constructed and transfected into DF-1 cells. The recombinant vector pCDH-CMV-bmp4-EF1-copGFP was constructed by homologous recombination technique, transfected into DF-1 cells, and the expression of bmp4 was detected by RT-qPCR. Three knockout target sites (gRNAs) were designed based on the bmp4 CDS sequence, and PGMLV-GM1 was inserted to construct the recombinant vectors bmp4-sgRNA1, bmp4-sgRNA2 and bmp4-sgRNA3. Cas9 vector was co-transfected into DF-1 cells with bmp4-sgRNA vector, and knockout vector activity and knockout efficiency were detected by T7E1 digestion and TA cloning assay. The full length of bmp4 CDS was 1546 bp, and the pCDH-CMV-bmp4-EF1-copGFP vector was successfully constructed. After transfected into DF-1 cells, it showed green fluorescence expression (25.17%±0.007). The results of RT-qPCR showed that bmp4 was significantly up-regulated in DF-1, after being transfected with pCDH-CMV-bmp4-EF1-copGFP compared with the control group (P<0.01). Three gRNA vectors were successfully constructed. The knockdown activity was detected by T7E1 digestion after transfection of DF-1 cells. The results of TA cloning showed that the knockout efficiencies were 6.25%, 12.5% and 18.75%, respectively. The results indicated that chicken bmp4 overexpression and knockout vector were constructed successfully, which could support the research of bmp4 function verification and mechanism analysis. Related Articles | Metrics

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Studies on genes and protein activities of extracellular superoxide dismutase homologs in Pacific oysters (Crassostrea gigas) #br# MAO Xiao-wei, XUE Qing-gang, XIA Qi-feng, LI Deng-feng, LIN Zhi-hua 2020, 37 (3):  35.  doi: 10.3969/j.issn.2095-1736.2020.03.035 Abstract ( 287 )   PDF   Save Superoxide dismutase (SOD) plays a role as an antioxidant by scavenging superoxide produced in organisms. Extracellular superoxide dismutase (Ec-SOD) is an important antioxidant protein in extracellular environments. However, Ec-SOD homologs in oyster plasma have been characterized to lack the SOD activity. In this study, 3 sequences that shared high similarity with dominin of the eastern oyster (Crassostrea virginica) were identified in the genome of the Pacific oyster (Crassostrea gigas) and verified by RT-PCR as real genes. The predicted mature peptide of Cg-dominin2, one of the 3 genes, was recombinantly expressed in Escherichia coli. The total antioxidant activity of the recombinant Cg-dominin 2 protein was equivalent to 0.0525 mmol/g Trolox. After challenges by zinc and cadmium ions, the relative expression of Cg-dominin2 gene was measured using qRT-PCR to be up-regulated significantly and the correspondent metal content of the oyster plasma proteins was also increased. These results suggested that the Ec-SOD homologs might play a role in defending against heavy metal stress in oysters by participating in metal metabolism. Related Articles | Metrics

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Identification of microRNAs in Gracilaria changii and phylogenetic evolution of their gene families SONG Wei, YIN Xu-gang, HE Zhong-shi, WANG Wei, FAN Yue, GAO Fan 2020, 37 (3):  40.  doi: 10.3969/j.issn.2095-1736.2020.03.040 Abstract ( 189 )   PDF   Save As an important representative of Gracilaria, Gracilaria changii has valuable industrial value. With deterioration of the global marine environment, Gracilaria resources are in urgent need for further development and utilization. As an important regulator of gene post-transcriptional expression, microRNA (miRNA) plays a key role in regulating plant tissues and organs, physiological status, stress response and so on. However, there have been no any reports about G. changii miRNA (gch-miRNA) in the world, which greatly hampers the further study on non-coding RNA and its function. Based on bioinformatics, six gch-miRNAs were predicted by sequences alignment analysis of gch-ESTs and miRNAs in miRBase databases in this study. The sequences and structural features showed that our prediction results were reliable. Moreover, the results of sequences alignment analysis showed that gch-miRNAs had both conservative and polymorphic features among other plants. Furthermore, gch-miRNAs validation showed our identifying results are high confidence by stem-loop qRT-PCR. The phylogenetic results of some miRNA families with NJ algorithm revealed the gch-miRNAs located in the outer group of the phylogenetic tree and the evolutionary rate of miRNAs is not synchronized with their corresponding host species. Eight potential gch-miRNA targets had been screened by TargetFinder software, and cleavage is the main inhibition type of miRNA(target in G. changii thorough psRNATarget analysis). This research firstly reported the miRNAs and their putative targets in G. changii, and comprehensively analyzed the phylogenetic relationship of some miRNAs across different plant species. The researching results lay a foundation for the non-coding RNA research on G. changii, as well as the molecular breeding of high-quality Gracilaria species and the development and utilization of them in future. Related Articles | Metrics

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Comparative analysis of intrinsic disordered proteins predictors based on random sequences #br# DONG Qing-qing, ZHAO Ya-wei, YUAN Zeng, YU Jia-feng, WANG Fang-hua, TANG Hu-cheng 2020, 37 (3):  46.  doi: 10.3969/j.issn.2095-1736.2020.03.046 Abstract ( 164 )   PDF   Save tIntrinsic disordered proteins (IDPs) are proteins that lack fixed 3D structures but perform important biological functions. Comprehensive evaluation of the prediction results of different IDPs predictors is basically important for IDPs studies.In this study, two kinds of IDPs predictors with different theoretical basis were analyzed based on randomly produced protein sequences. The results indicated that the prediction results are different to different degree based on each kind of predictor.It is difficult to decide which is better for IDPs prediction programs, more efficient prediction methods should be proposed in future studies. Related Articles | Metrics

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Antianxiety effect of long-term treatment of Konjac mannan-oligosaccharides in mice WANG Xiao-qin, MA Yan-fang, TANG Xiang-hua, WANG Gong-wu 2020, 37 (3):  50.  doi: 10.3969/j.issn.2095-1736.2020.03.050 Abstract ( 180 )   PDF   Save Konjac mannan-oligosaccharides (KMOS) are a kind of important prebiotic. In present study, in order to investigate the effects of KMOS on emotional neural behaviors and the hippocampal neuron density in mice, we divided 50 adult male KM mice into 5 groups， and fed them with KMOS of 0 (control), 650, 1400, 3000 and 6500 mg/kg (i.g.) for consecutive 30 d, and then the mice were tested in the open-field, elevated plus maze (EPM) and forced swimming tasks. At last, their brains were sectioned and stained for hippocampal neuron density counting. Results showed that, the ventral hippocampal CA3 neuron density of KMOS group (6500 mg/kg) increased significantly (P<0.01, vs. control) and positively correlated with the doses (P<0.01). In open field, the rearing times and center entries of KMOS groups (1400 and 3000 mg/kg) significantly increased (P<0.01, vs. control). In EPM, the open arms entries of mice in 3000 mg/kg group also increased (P<0.01, vs. control). The results suggested that long-term treatment of mice in KMOS would be involved in neuron survival in hippocampal CA3 subarea and have anxiolytic effect in mice. Related Articles | Metrics

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Protective effect of Spirulina polysaccharide on liver, heart, kidney and thymusliver oxidative injury induced by acute alcoholism in mice #br# CHEN Shuai-hang, MA Hao-tian, LI Run-zhi, GE Bao-sheng, CUI Hong-li 2020, 37 (3):  54.  doi: 10.3969/j.issn.2095-1736.2020.03.054 Abstract ( 308 )   PDF   Save Consuming large quantities of alcohol over a short time can lead to acute alcoholism, which usually causes gastrorrhagia and cardiovascular disease. These diseases have become the most common clinical symptoms. Recently, synthetic drugs used to treat acute alcoholism have side effects on the body. Therefore, it is urgent to discover some natural active substances that can replace synthetic drugs. The goal of this article is to study the protective effect of polysaccharide from spirulina on acute alcoholism. Thirty mice were randomly divided into control (blank), model (alcohol injury), positive control, treatments (low PSP and high PSP dose). The low PSP and high PSP dose treatments were continuously taking polysaccharide from spirulina for 21 days (1 time/d). The positive control was given a continuous bifendate pills for 21 days (1 time/d). The blank control and alcohol injury were given solvent (water) for 21 days (1 time/d). After 22 days, except for the control group, others were given 50% alcohol (12 mL/kg) by means of intragastric. All groups mice were killed 12 hours later, and then the hearts, kidneys, thymus and livers were dissected from mice. The tissues were ground and then the supernatants were collected. The activities of catalase (CAT), glutathione (GSH), malondialdehyde (MDA), alanine aminotransferase (ALT) and aspartic aminotransferase (AST) were analyzed. The results showed that compared with the model group, the contents of CAT and GSH in treatments increased significantly and the content of MDA decreased dramatically. In liver tissue, the contents of ALT and AST decreased distinctly and returned to the level of positive control group. The above results would provide a scientific basis for the treatment of oxidative damage in the kidney, heart, liver and thymus caused by acute alcoholism. Related Articles | Metrics

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Inhibition effects of arsenic trioxide on gonad tumorous proliferation of the nematode Caenorhabditis elegans YU Min, LIU Tian-jiao, YOU Mu, WANG Shun-chang 2020, 37 (3):  58.  doi: 10.3969/j.issn.2095-1736.2020.03.058 Abstract ( 224 )   PDF   Save Inorganic arsenic has been shown as an environmental carcinogen, exposure to arsenic can resulted in high incidence of multiple diseases including cancer. Paradoxically, several arsenic compounds such as arsenic trioxide have been used effectively to treat promyelocytic leukaemia and other solid tumors. It has been shown that the tumor suppressor p53 plays important roles in cancer therapy. However, the roles of p53 on the treatment effects of ATO are still uncertain. The nematode Caenorhabditis elegans has been shown as a good model for studying the regulative effects of p53 on DNA damage and cell cycle because it has only a single p53-like gene cep-1. In the present study, the therapeutic effects of ATO on the gonad tumorous growth were studied using gld-1(q485) mutated worms. The results presented here indicated that ATO treatment extended the lifespan of gld-1(q485) mutants and inhibited tumorous growth of the gonad. ATO treatment induced cell cycle arrest in a dose- and time-dependent manner. It was also found that ATO treatment resulted in germline apoptosis in worms of N2 and gld-1(q485) mutants. By using worm strains carrying mutated cep-1 alleles plus RNAi double gene knockout, it was found that ATO exerted its antitumor activity on gld-1(q485) mutants through the induction of cell cycle arrest in a CEP-1/p53-independent manner and germline apoptosis in a CEP-1/p53-dependent manner. Related Articles | Metrics

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Estimation features of nitrous oxide in Phragmites australis-dominated wetland of Baiyangdian Lake by DNDC model FENG Zhao, QIAO Sheng-chao, NI Shao-qiang, ZHANG Jie, YU Chao-qing 2020, 37 (3):  62.  doi: 10.3969/j.issn.2095-1736.2020.03.062 Abstract ( 190 )   PDF   Save Wetland is a major source of nitrous oxide. Climate changes can influence N2O emission. To explore its emission′s characteristics of wetland ecosystem during the growth of Phragmites australis from 2018 to 2100, three RCP (Representative concentration pathway) scenarios in six models selected from IPCC (International Panel on Climate Change) were used by DNDC (Denitrification decomposition) model. The results showed that multipoint historical position experiments present a clear relationship between observed values and simulated ones, and correlation coefficient r is 0.723, while determination coefficient (R2) is 0.522 with RMS(Root mean square error) of 76.75 μg/m2 · d. No obvious and regular fluctuations are found in the condition of RCP2.6 and RCP4.5 scenarios, whose annual nitrous oxide emission range from 0.69-23.84 and 0.59-31.92 simulated by DNDC in the future. Compared to RCP 8.5 scenario, N2O emission increases significantly in both CanEMS2 and MIROC5 with year total emission from 0.49-38.82, respectively. N2O emission does not show a significant increasing or decreasing trend in the three sections divided in the period of 2008-2100. Under multi-modes and multi-scenarios, the N2O emission has no significant periodic changes but it is affected obviously by precipitation and temperature. The maximum N2O emission is likely to occur during the year with moderate temperature and abundant precipitation. Related Articles | Metrics

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Epigenetic diversity and structure of Phragmites australis in heterogeneous habitats in the Songnen Meadow QIU Tian 2020, 37 (3):  68.  doi: 10.3969/j.issn.2095-1736.2020.03.068 Abstract ( 171 )   PDF   Save Epigenetics has not only become a popular field in biology, but also undergone rapid development in plant ecology due to an increasing number of studies. However, few assessments of the spectra and structures of epigenetic variation have been reported on wild plant populations. The epigenetic diversity and structure of naturally occurring Phragmites australis populations, occupying two different habitats on a small spatial scale in the Songnen Meadow in northeastern China, were investigated by assessing methylation-sensitive amplified polymorphisms (MSAPs) through fluorescent capillary detection. The habitat 1( i.e. H1) was a low-lying alkalized meadow with seasonal rainfall and pH 8-8.5. The habitat 2 ( i.e. H2) did not accumulate rainwater with pH higher than 10 as an alkaline patch. Among the methylation-susceptible loci (MSL), 71.09%-88.38% of which were polymorphic. At the intra-habitat level, 82.75% and 79.93% of the loci observed from H1 and H2 were polymorphic, with I indiex of 0.3712 and 0.3580, respectively, showing higher values in H1 than in H2. The AMOVA-based MSAP analysis showed significant epigenetic differentiation between habitats（P＜0.01）. Epigenetic variations of individuals would provide more phenotypes and evolutionary flexibility. Remarkable epigenetic differentiation between habitats would underly the basis of different phenotypes of P. australis. In a word, this study attempted to explore the ecological adaptation of reeds in different habitats on a small scale, which would provide guidelines when seeking to understand processes that drive phenotypic evolution and adaptation. Related Articles | Metrics

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The inhibition effect of two common limnetic submerged macrophytes on Scenedesmus obliquus under co-culture condition #br# MA Hao-tian, ZHANG Fei, ZHANG Hong-jiang, SHI Fei-fei, CUI Hong-li, LI Run-zhi 2020, 37 (3):  72.  doi: 10.3969/j.issn.2095-1736.2020.03.072 Abstract ( 244 )   PDF   Save Submerged macrophytes and microalgae are important components of the aquatic ecosystem. There may be a competition between them, and submerged plants can often take advantage by allelopathic effects. In order to explore the allelopathic inhibition effect of submerged plants on freshwater microalgae, Scenedesmus obliquus was selected as the test organism, the growth and physiology properties were detected under the Potamogeton Malaianus, Ceratophyllum demersum and their mixed culture condition. The results showed that under the condition of co-culture with submerged plants, the growth of S. obliquus was significantly inhibited, and the chlorophyll content decreased, the chlorophyll autofluorescence changed, which probed the damage of microalgal photosynthetic system, and the increase of malondialdehyde and antioxidant enzymes activities could illustrate the damage of the antioxidant system. All the results proved that both submerged plants and their mixed cultures had strong algae-inhibiting effects, which could lead to a series of damage to algae cells. The above research results would provide a scientific basis for the investigation of the mechanism of allelopathic inhibition, the analysis of related phenotypes, and the ecological relationship between submerged plants and microalgae. Related Articles | Metrics

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Enrichment of heavy metals and health risk assessment of plants in Tongling mining area WANG Juan, LI Yu-cheng, HUANG Xin-xin, ZHOU Shao-qi, ZHANG Xue-sheng, GAO Yi, LIU Bing-xiang 2020, 37 (3):  76.  doi: 10.3969/j.issn.2095-1736.2020.03.076 Abstract ( 298 )   PDF   Save Twenty-seven rhizospheric soils, 14 main crops, including rice (Oryza sativa) and maize (Zea mays L.), 10 garden vegetables, including eggplant (Solanum melongena L.), chili (Capsicum annuum L.) and greenbean (Vigna unguiculata), and 8 medicinal plant Fengdan (Paeonia ostii), were collected in Tongling mining area. The aim was to analyze five heavy metals (Cu, Zn, Cd, Pb and Cr) enrichment and transport characteristics and to assess their health risks. The results showed that the mining activities had caused the enrichment of 5 metal elements in rhizospheric soils. Cu, Zn, Cd and Pb were seriously polluted. The pollution degrees showed the trend of Cd＞Cu＞Zn＞Pb＞Cr. The accumulation of heavy metals in the fruits of the investigated plants and chili in the vegetables was the lowest. Cr and Cd had strong migration ability and were easily absorbed and accumulated by plants. Heavy metal contents, daily estimated intake (EDI) and target hazard quotient (THQ) jointly indicated that Cr, Cd and Pb were mainly enriched in rice; Cr, Cd and Pb were mainly accumulated in maize, vegetables and Fengdan. These elements were all of great health risks to local populations. Related Articles | Metrics

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Effects of ecological comprehensive planting and breeding patterns on soil physical and chemical properties and humus in paddy fields YU Jing-wei, HUANG Wei, LI Yu-cheng, WANG Ning, ZHANG Xue-sheng 2020, 37 (3):  81.  doi: 10.3969/j.issn.2095-1736.2020.03.081 Abstract ( 231 )   PDF   Save On the basis of the chemical fertilizer reduction, this study explored the effects of paddy integrated ecological system on the physical and chemical properties and humus of paddy soils. Four experimental fields were selected around Chaohu lake basin. The large-scale integrated ecological model (Early rice crayfish rotation, later rice-duck mutalism) of paddy field was compared with conventional cultivation models (idle, single-season rice cultivation and double-season rice cultivation), with a cycle of 2 years. The results showed that there were differences in soil bulk density and total porosity components in the four types of planting models（P<0.05）. Compared to the conventional planting models, PES was effective in soil texture, which improved sand differentiation rate and soil total porosity, and reduced soil bulk density. In PES model, the soil fertility factors increased significantly. The activity and degree of humus were improved. At the same time, the degree of condensation and molecular complexity of humic acid in soils decreased significantly. The PES model has improved the physical structure and fertilizer retention capacity of paddy fields and increased the economic income of planting, which is worth popularizing. Related Articles | Metrics

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Isolation of room temperature cellulose-degrading bacteria and construction of degrading consortia MENG Jian-yu, JI Jin-hua, GUO Hui-qin, TAO Yu, FENG Fu-ying, ZHAO Hong-bin 2020, 37 (3):  86.  doi: 10.3969/j.issn.2095-1736.2020.03.086 Abstract ( 260 )   PDF   Save The room temperature cellulose-degrading bacteria were screened from the forest soil of Hulunbeier city. The strains of high enzyme activity were separately mixed to construct room temperature cellulose-degrading consortia, and filter paper degradation abilities were measured, using three different substrates (avicel, CMC and D-salicin) as carbon source respectively to isolate cellulose-degrading bacteria at 25 ℃. The 16S rDNA sequence was used for identification and phylogenetic analysis of strains, and DNS method was used to determine the enzyme activity of exogenous glucanase, endogenous glucanase and β-glucanase of the isolated strains, as well as the filter paper enzyme activity of the cellulose-degrading consortia. As a result, 258 room temperature cellulose-degrading bacteria were isolated. Among them, the bacteria isolated with avicel as substrates belong to five classes, and the bacteria isolated with CMC or D-salicin as substrates belong to six classes. The first dominant class was γ-Proteobacteria, and the second dominant class was α-Proteobacteria. The first dominant genera was Pseudomonas, followed by Rhizobium, Oerskasty, Bacillus and Lelliottia. In 27 cellulose-degrading consortia, C11′3″, C32′2″ and C33′2″ had the highest filter paper enzyme activity, and the enzyme activity was 200.60, 200.05 and 208.79 U/mL, respectively, which is 6-10 times more than the filter paper enzyme activity of single strain in its consortia. The degradation rate of filter paper was 26.8%, 22% and 32.4%, respectively. In conclusion, C11′3″, C32′2″ and C33′2″ are optimal flora combination of cellulose-degrading consortia, which have higher application potential. Related Articles | Metrics

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Research advances in plant endophytes Bacillaceae GONG Guo-li, WANG Liang, WANG Xu-yang, ZHANG Di, WU Mi, TIAN Lu 2020, 37 (3):  91.  doi: 10.3969/j.issn.2095-1736.2020.03.091 Abstract ( 403 )   PDF   Save Bacillaceae is an important part of plant ecology, which is widely distributed and diverse, and plays an extensive and necessary role in the growth cycle of plants by promoting growth, resisting diseases and pests, and resisting adversity. Endophytic Bacillaceae and its products not only have great application value in the field of agricultural biological control, but also provide a new feasible idea for solving the problems of new drug research and development, environmental restoration and so on. In this paper, the diversity of endophytic Bacillaceae, the function of endophytic Bacillaceae to host plants by secreting or inducing effector molecules such as growth hormone, antimicrobial proteins and lipopeptide, enzymes, etc. were reviewed. In addition, the diversity of endophytic Bacillaceae and the functional mechanism of effector molecules were summarized and prospected. Related Articles | Metrics

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Recent advances on the hypoglycemic effect and mechanism of dietary nutrients in type 2 diabetes mellitus XU Fei, CHEN Lin-jun, NIE Zhi-yan , NA Li-xin 2020, 37 (3):  96.  doi: 10.3969/j.issn.2095-1736.2020.03.096 Abstract ( 269 )   PDF   Save Type 2 diabetes mellitus (T2DM) is a complex disease resulting from the interaction of genetic, epigenetic, environmental, and lifestyle factors. The morbidity and mortality rates of T2DM are increasing year by year. The pathogenesis of DM is not completely understood, but more and more studies show that T2DM is related to genetic susceptibility, dietary pattern and nutrient-gene interactions. However, people have known little about the interaction and its mechanism between dietary factors and genes. Nutrigenomics research can not only help us better understand the hypoglycemic effect and mechanism of dietary nutrients in T2DM, but also help to establish personalized diet intervention methods and nutritional health care methods, so that the results of nutritional research can be more effectively applied to the prevention of chronic diseases to achieve the goal of promoting human health. In this paper, the research progress of the hypoglycemic effect and mechanism of dietary nutrients (polyphenols, dietary fat, vitamins and amino acids) in T2DM was reviewed. Related Articles | Metrics

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Optimization of the extraction process and analysis of Dipsacu asperides root protein #br# JIN Hai-jun, JIANG Ting-hong, WANG Hai-xia, SHEN Xu-feng, YU Hua, ZHU Shan, ZHANG Jia 2020, 37 (3):  102.  doi: 10.3969/j.issn.2095-1736.2020.03.102 Abstract ( 225 )   PDF   Save In order to promote the proteomic investigation of Dipsacu asperides roots, it is necessary to analyze the protein extraction process and SDS-PAGE gel electrophoresis patterns of the roots. In this study, Tris-HCl method was used to extract the protein, and the differential bands of the root proteins from different producing areas were analyzed by SDS-PAGE gel electrophoresis. The extraction process was optimized by response surface. The results showed that the protein content of D. asperides roots in Hubei was the highest, followed by Guizhou and Yunnan. The gray value analysis of gel electrophoresis showed that two protein bands at 24 ku and 34 ku were significant differences in different habitats (Yunnan, Hubei and Guizhou) (P<0.05). Response surface methodology (RSM) was used to optimize the extraction process of D. asperides root protein, and the optimum extraction conditions were as follows: extraction temperature, 16.84 ℃; extraction time, 0.84 h; pH value, 6.58; solid-liquid ratio, 1∶ 1.64. The screened differential protein bands and the optimized extraction technology of D. asperides root protein can provide basic data for further understanding the molecular mechanism of the quality difference of D. asperides from different producing areas. Related Articles | Metrics

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Optimization of biological preparation of human serum albumin CAI Yan-fei, CHEN Yun, JIN Jian 2020, 37 (3):  106.  doi: 10.3969/j.issn.2095-1736.2020.03.106 Abstract ( 632 )   PDF   Save The human serum albumin was prepared by the expression system of CHO cells, which provided a basis for the preparation and function research of HSA and its protein-fusion drugs. Firstly, the recombinant expression plasmid pMH3-HSA was constructed. Then the positive monoclones were screened by finite dilution and Dot Blot. High density amplification culture by 5 L disposable bioreactor was used after suspension domestication, the serum-free medium B001 and F001 were used as basic and flow media, respectively, the fermentation parameters were 55 r/min, DO 20%-40%, pH 6.8-7.4, and Tm 37 ℃. The cell growth status and expression of target protein HSA during fermentation were detected by cell counts, SDS-PAGE, Elisa and Western Blot, and compared with the HSA protein prepared in the Pichia expression system. The results showed that the highly expressed monoclonal CHO cell line containing exogenous HSA gene was successfully constructed and screened, the maximum yield of HSA reached 180 μg /mL in 5 L fed batch disposable bioreacter, and the product of CHO expression system had almost no degradation. In summary, this study indicated that HSA was more suitable for expression in the CHO system, and its advantage of protein integrity was particularly obvious in this system. Related Articles | Metrics

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Simultaneous quantification of free amino acids in Dendrobium huoshanense by a precolumn derivatization-HPCE technology CHEN Nai-dong, ZHU Shai , WANG Rong-hua, XU Wen-dong, SHI Min-zhu 2020, 37 (3):  110.  doi: 10.3969/j.issn.2095-1736.2020.03.110 Abstract ( 263 )   PDF   Save As the substrates of Maillard reaction, free amino acids were the key matters which may greatly affect the chemical compositions in traditional Chinese medicines and the flavored constituents in food. Dendrobium husohansense, a rare and endangered medicinal plant, has long been used in clinical as its processed dry stems due to the great difficulty in the preservation of its fresh stems. Thus, it is essential to assay the free amino acids for the investigation of the processing mechanism of D. huoshanese. In this study, we established an efficient high performance capillary electrophoresis (HPCE) method to qualify and quantify the free amino acids in D. huoshanense. The results showed that the optimal HPCE method was as follows: detection wavelength at 254 nm, separation voltage of 10 kV with effective length of uncoated standard fused silica capillary 55 cm. The running buffer was 60 mmol/L Na2B4O7-Na2HPO4 with pH value of 11.0. Under these conditions, all the 12 investigated amino acids could be separated in 60 min with the R2 values of all the calibrations for the 12 amino acids more than 0.99 and recovery within 90.17%-110.25% (RSD≤5%, n=3). The total contents of total amino acids in the root, stem and leaf in D. huoshanense were 1.8958, 2.0736 and 2.5954 mg/g, respectively. These conformed the established method in our experiments is accurate, reliable and reproducible, and can be used to qualify and quantify the free amino acids in D. huoshanense. Related Articles | Metrics

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Thoughts and practice about cell biology teaching CUI Ji-hong, LI Jun-lin, HUANG Xuan, TAN Zeng-qi, FU Ai-gen, CHEN Fu-lin 2020, 37 (3):  115.  doi: 10.3969/j.issn.2095-1736.2020.03.115 Abstract ( 319 )   PDF   Save In the new period, the cultivation of innovative entrepreneurial talents requires higher requirements for higher undergraduate education. Curriculum teaching is the core of personnel training. In this paper, the authors consider and practice how to build "Gold Course" of cell biology teaching according to three requirements of the construction of "Gold Course": teacher-student interaction, process of concern and evaluation system. On the basis of traditional cognition such as "teachers for the guider of teaching activities" and "Students for the subject of learning", they emphasize the interaction between teachers and students. In terms of teaching activities and evaluation system, the emphasis is placed on the process of guiding, cultivating and evaluating students′ independent learning, cooperation, thinking, mastering basic skills and promoting the progress of open issues from the 3 stages of preview, in-class and after class. On the premise of the above, the goal is to achieve personnel training. Related Articles | Metrics

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Utilization of Adobe Photoshop in drawing plant structural picture MA San-mei, WANG Yong-fei 2020, 37 (3):  118.  doi: 10.3969/j.issn.2095-1736.2020.03.118 Abstract ( 342 )   PDF   Save Adobe Photoshop is a very excellent software for image processing. Its toolbox is vivid, easy to distinguish and easy to use. The brush in the toolbox can be used to draw on a computer screen. Graphics tablet is usually composed of a board and a pressure pen, which is a kind of computer input equipment. When a curve is drawn on the board with a pressure pen, then this curve appears on the computer screen. Combining Adobe Photoshop with a graphics tablet, drawing on a computer screen and drawing with a pencil has the same sensation. The pictures drawn by Adobe Photoshop and graphics tablet are colorful, easy to modify, copy, share and have high resolution. Using Adobe Photoshop and graphics tablet to draw the structure of plant slices on computer screen has many advantages. In this paper, we mainly introduce the basic skills of drawing the structure of water lettuce root using Adobe Photoshop and graphics tablet. The skill of drawing plant structure can be learned through the process of drawing root structure of water lettuce, and then you can draw colorful plant structure of other plants. Related Articles | Metrics

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Application of DNA shuffling of plant resistance genes in genetics experimental teaching ZHANG Yuan-li, PANG Yan-jun 2020, 37 (3):  121.  doi: 10.3969/j.issn.2095-1736.2020.03.121 Abstract ( 280 )   PDF   Save Genetics experiment is a core experiment course in life science, medicine, agriculture and other related fields. In order to stimulate students′ interest in learning genetics experiment, some genetics experiments about scientific research were designed in this class. In this comprehensive experiment, plant resistance genes were synthesized by DNA shuffling technology. Gene fragments were cloned from different rice varieties, and new genes were obtained by artificial evolution using the DNA shuffling technology. Through these comprehensive experiments, students can understand the basic operation of molecular genetics and the basic phenomena and laws of genetics at the molecular level. Compared with the traditional genetic experiment, students can also improve the ability to carry out scientific research. This will provide a platform for the cultivation of innovative talents. Related Articles | Metrics

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Reform and practice of zoology curriculum guided by cultivating innovative talents WANG Dan-li, ZHAO Chun-pu, LI Ge, WANG Ya-jun, XU Shan-liang 2020, 37 (3):  124.  doi: 10.3969/j.issn.2095-1736.2020.03.124 Abstract ( 298 )   PDF   Save The university′s zoology course is a compulsory course for the basic education platform of biology, which is very important for follow-up professional learning. Based on the cultivation of innovative undergraduate students in aquaculture, in view of the thoughts and learning characteristics of the "After 95" and "After 00" generation, we analyzed the existing problems in current zoology classroom teaching, and put forward the countermeasures and suggestions for teaching reform from the following 4 aspects: extended thinking training before, during and after class guided by "problem-led"; holographic experience of "physical specimen -digital model - two-dimensional information" by multidimensional interaction; student research practice and teaching software development with the goal of “self-participation” ; "scientific evaluation" as the result of student self-evaluation system innovation. This paper summarized the reform and practical experience of zoology curriculum teaching in recent years, in order to further enhance the effectiveness of zoology curriculum teaching, and improve students′ expanding thinking and innovative ability, so as to lay a solid foundation for cultivating innovative talents and outstanding talents in marine science. Related Articles | Metrics

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Practice of virus-induced gene silencing technology in general biology experiment teaching #br# ZHU Feng, JI Zhao-lin, QIAN Kun 2020, 37 (3):  127.  doi: 10.3969/j.issn.2095-1736.2020.03.127 Abstract ( 232 )   PDF   Save General biology experiment has the characteristics of strong practicality and maneuverability, and it plays an important role in the cultivation of biology professionals. In order to discipline and cultivate students′ experimental operation ability, innovative consciousness and innovative ability in experimental teaching and improve the teaching quality of general biology for biology major in local colleges and universities, the author introduced virus-induced gene silencing (VIGS) technology, a hot topic in current biology research, into teaching experiments. Through this experimental teaching, students′ interests are obviously improved, their independent working ability and innovative thinking ability are cultivated, so that students can initially form a certain scientific research ability and innovative ability. Related Articles | Metrics