Table of Content

18 August 2016, Volume 33 Issue 4 Previous Issue    Next Issue

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Cloning and prokaryotic expression analysis of Unigene A09801from Cladosporium cladosporioides MD2 XI Xiao-yuan, ZHANG Peng, SONG Fa-jun, YU Peng-fei, WANG Qi, MENG Yan-yan 2016, 33 (4):  1.  Abstract ( 245 )   PDF   Save Molecular cloning and functional research on genes related to fungal taxol biosynthesis is a key for the genetic engineering improvement of taxol-producing fungi. In this study, the full length sequences (1674 bp) of DNA and cDNA of Unigene A09801 were cloned from the taxol-producing fungus, Cladosporium cladosporioides MD2, respectively. Unigene A09801 sequence had no introns. Unigene A09801 encoding protein contained 557 amino acids, had 56% identity with GMC (glucose-methanol-choline) oxidoreductase, possessed 61 291.8 u molecular weight and 5.47 isoelectric point value, and had no trans-membrane structure and signal peptide. Unigene A09801 existed in the fungal genome with a single copy. The expression levels of Unigene A09801 responded to Methyl Jasmonate were showed to decrease firstly, increase subsequently and then decrease, and the highest expression level appeared at 24 h. The prokaryotic expression vector of Unigene A09801 was constructed, and the optimized expression conditions of Unigene A09801 in E. coli BL 21 were: induced at 32℃ for 4 h, with 1.2 mmol/L TPTG. These results would lay a foundation for further in vitro analyzing catalytic function of Unigene A09801 encoding protein. Related Articles | Metrics

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Molecular cloning, sequence analysis and expression analysis of ApoC-Ⅰgene from ayu (Plecoglossus altivelis) exposed to cadmium CHEN Ya-ping, DING Fei-fei, LI Chang-hong, MO Li-bin, CHEN Jiong 2016, 33 (4):  5.  Abstract ( 260 )   PDF   Save ApoC-Ⅰ(Apolipoprotein C-Ⅰ) is the smallest member of the apolipoprotein family, which plays important roles in lipid uptake, secretion and transport. Here, we obtained the full-length cDNA sequence of ayu(Plecoglossus altivelis) ApoC-I gene (PaApoC-Ⅰ) through random sequencing of cDNA library. The cDNA sequence of PaApoC-Ⅰis 574 nucleotides, consisting of a 267 bp open reading frame that encodes a protein of 88 amino acids with a deduced molecular mass of 10.08 ku. The N-terminus of PaApoC-Ⅰcontains a predicted signal peptide of 20 amino acids. Multiple sequence alignment of complete amino acid sequences showed that PaApoC-Ⅰshared the highest amino acid sequence identity (67.0%) with that of rainbow trout (Oncorhynchus mykiss). Phylogenetic tree analysis revealed that ApoC-Ⅰfrom mammals, reptiles, amphibians and fish formed different clusters, and PaApoC-Ⅰfalled into the fish ApoC-Ⅰcluster and was most closely related to that of rainbow trout. Genome structure analysis showed that PaApoC-Ⅰcontained three exons and two introns. Quantitative real-time PCR (qRT-PCR) analysis showed that PaApoC-ⅠmRNA had the highest expressional level in ayu liver. Upon cadmium (Cd) exposure, PaApoC-I transcripts were significantly up-regulated in ayu liver, spleen, kidney and intestine, and were significantly down-regulated in ayu gill. In summary, our present study demonstrated that a tight relationship between the PaApoC-Ⅰ expression and Cd stress, and it can be a potential biomarker for Cd exposure in aquatic organisms. The results would provide the basic data for further studying the function of fish ApoC-Ⅰ and its mechanism in fish response to heavy metal stress. Related Articles | Metrics

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Molecular characteristics and expression analysis of Kazal-type serines proteinase inhibitor (KSPI )gene from Hyriopsis cumingii XU Long-wei,WANG Qin, WANG Gui-ling, LI Jia-le 2016, 33 (4):  10.  Abstract ( 282 )   PDF   Save Kazal-type serines proteinase inhibitor (KSPI) is very important in the immune reaction of biological body, but the study of KSPI in Hyriopsis cumingii has been rarely reported. In this study, the full length of cDNA sequence of Kazal-type serines proteinase inhibitor(KSPI) gene (accession number: KT901291) of H.cumingii was cloned. It was 1029 bp in length, containing 5′and 3′-UTRs parts of 61 bp and 206 bp, and an ORF of 762 bp encoding 253 amino acids, and the molecular weight was 27397.5 u. The amino acid sequence contains five Kazal domain structures, and the result of homology analysis showed that it has a few of similarity with other species, suggesting that this gene belongs to the typical KSPI gene. Real-time quantitative PCR showed that KSPI gene expressed in a wide range of tissues including the adductor, foot, liver, blood, mantle, gill and gonad, and the highest expression was in mantle, on the contrary, the lowest in liver. After being infected with Aeromonas hydrophila， the expression of KSPI gene had significant increasing in 7 tissues compared with the control group. The results suggested that KSPI gene is very important in the immune reaction of Hyriopsis cumingii. Related Articles | Metrics

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Constructing and applying recombinant plasmid of clock and bmal1 3′UTR WANG Yan-li, LV Ke, CHEN Hai-long, JI Guo-hua, WANG Ting-mei 2016, 33 (4):  15.  Abstract ( 279 )   PDF   Save tTo investigate the effects of miRNAs on the post-transcriptional regulation of circadian genes, we first constructed the clock and bmal1 3′UTR recombinant plasmid. The 3′UTR sequence of mouse clock and bmal1 genes searched from the NCBI was awplified, and then cloned into the pGL3-promoter plasmid. Plasmid DNA was subsequently sequenced to ensure the authenticity and direction of the inserted clock and bmal1 3′UTR. Next, the miRNAs modulation clock and bmal1 expression was analyzed by detecting the luciferase activity. The miRNAs targeting clock, bmal1 gene 3′UTR was predicted by TargetScan. The luciferase activity was analyzed in 293T cells which were cotransfection with recombinant plasmid and miRNA mimic by LP2000. Result showed that the sequences analyzed by RVP4 primer were inversely complementary to the sequences in NCBI. The mouse clock and bmal1 3′UTR recombinant plasmids were successfully constructed, which may lay a foundation for further study of post-transcriptional regulation mechanism of circadian genes. Also the results showed that the luciferase activity of clock and bmal1 3′UTR recombinant plasmid was reduced 33% and 45% by miR-199a-5p（P<0.05）and miR-142-3p(P<0.01)， respectively. The activity of clock and bmal1 gene was modulated by miR-199a-5p and miR-142-3p, respectively. Related Articles | Metrics

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The heterologous expression and application of Endo-β-N-acetyglucosaminidase H from Streptomyces avermitilis in Bacillus subtilis LIANG Biao, GUO Jia,GUAN Feng 2016, 33 (4):  20.  Abstract ( 487 )   PDF   Save Endo-β-N-acetylglucosaminidase H (Endo H) is a glycohydrolase which cleaves the β-1,4-glycosidic bonds of the N-acetylglucosamine core of oligosaccharides and leaves one N-acetylchitobiose attached to the asparagine residue of the glycoprotein. It can be used to identify the glycosylation sites. In this study, we cloned the endoH gene from the Streptomyces avermitilis, then subcloned into expression vector pMA0911.1. We expressed EndoH in Bacillus subtilis WB600. The fermentation broth was purified by hydrophobic chromatography and ionic exchange, and relatively purified EndoH was obtained. Utility of EndoH in digestion of RNaseB and identification of N-glycan structure in OVA indicated the potential of recombinant EndoH in research of N-glycan and glycomic. Related Articles | Metrics

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Isolation of alkaline pectinase-producing strain and the research on separation and purification of its pectinase TAN Xiu-hua 2016, 33 (4):  25.  Abstract ( 239 )   PDF   Save A bacterial strain producing high-activity and high-temperature alkaline pectinase was obtained from the soil and was named as AP-1. The characteristics and the separation and purification of the pectinase produced by AP-1 were preliminarily studied. The result of the study on pectinase activity showed that the optimal temperature and pH of the pectinase were 70℃ and 7.5, respectively. The experiment of stability showed that the enzyme had good resistance to alkali and was activated remarkably by Mg2+ under the same conditions. The separation and purification showed that the purification effect was better when the enzyme was treated by 80% saturation of ammonium sulfate solution than other, but the purification effect was the best when the enzyme was treated by ethanol. Related Articles | Metrics

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Analysis and comparison of biochemical composition in squid ink between wild and cultured Sepia pharaonis JIANG Mao-wang, JIANG Xia-min, GAO Xiao-lan,WANG Peng-shuai, RUAN Peng, HAN Qing-xi 2016, 33 (4):  29.  Abstract ( 308 )   PDF   Save This study was conducted to provide a basis for the research and development of the nutrient composition in squid ink between wild and cultured Sepia pharaonis. The moisture, crude ash, crude protein, crude fat, polysaccharide, amino acids, fatty acids and minerals were determined by national standard nutritional method. The results showed as follows: the moisture of wild and cultured Sepia pharaons squid ink was not significantly different (P>0.05), whereas other basic nutrient contents (dry weight basis) were significantly different (P<0.05). The content of polysaccharide was abundant and in wild squid ink (3.25%) with cultured squid ink (29.18%). 4 kinds of constant elements (Na, Ca, K and Mg) and 7 kinds of trace elements (Cu, Fe, Zn, Al, Mn, Sr and As) were determined, and 8 kinds of elements showed significant difference (P<0.05) with the exception of Mg, Mn and Al. The trace element (Zn) was high, with cultured squid ink (5.40±0.71)mg/g>with wild squid ink (3.89±0.19)mg/g. In conclusion, cultured Sepia pharaonis squid ink has higher nutritional value, medicinal and health care function, with promising further development. Related Articles | Metrics

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Comparison of blood parameters, antioxidant enzyme activity in liver and muscle of rainbow trout (Oncorhynchus mykiss) following exposure in freshwater and seawater GONG Jian-hua, GUO Chun-yang, TIAN Zhe, XU Shan-liang 2016, 33 (4):  34.  Abstract ( 265 )   PDF   Save Using Beckman electrolyte and automatic biochemical analyzer, we compared the blood index, and the activity of antioxidant enzyme in muscle and liver under different circumstance . It turned out that under sea and fresh water environment, the content of glutamic pyruvic transaminase（GPT）are significantly different（P<0.05）, so are the contents of aspartate transaminase（GOT）. The contents of fish in GDT and GOT in freshwater are higher than those of seawater, the values are （7.00±2.82）μ/ L and（44.00±7.43）μ/ L, respectively, which are 1.36 times and 3.35 times of the seawater group. The trace element potassium （K+）, sodium （Na+）, chlorine（Cl-）, calcium （Ca2+) and albumin（A）, triglycerides and globulin（G）in sea wafer fish are greater than those in freshwater group. Total cholesterol levels were markedly different（P <0.05）, the level of seawater group is greater than that of freshwater group. Hemoglobin, platelet hematocrit, number of red blood cells, platelet count of fishes in seawater are obviously higher than those in freshwater group, which decreased by 68.31%, 37.32%, 22.58% and 24.29%, respectively; The width of platelet distribution and red blood cell distribution and the hematocrit is apparently different（P<0.05）. Related Articles | Metrics

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Study on extrusomes of the hypotrichous ciliate Oxytricha granulifera (Ciliophora: Hypotrichida) TANG Wen-jun, FAN Xin-peng, NI Bing, GU Fu-kang 2016, 33 (4):  38.  Abstract ( 275 )   PDF   Save Ultrastructural observation showed that a great number of long elliptical extrusomes were distributed beneath the pellicle on the dorsal and ventral sides of a hypotrichous ciliate, Oxytricha granulifera. The mature extrusomes were filled with different electron dense fibrous materials at their posterior parts, and had hollow cavities at their anterior parts. The anterior end of the mature extrusomes was close to the pellicle, and distributed in longitudinal, diagonal or arc rows. During ejection, the anterior membrane of extrusomes fused with the plasma membrane. After extrusion, the extruded contents often remained in the surface of the cells. Considered that the extruded contents of extrusomes of O. granulifera always adhere to the surfaces of the cells and its morphology has no obvious difference from the resting extrusomes, it can be supposed that during ejection, the contents may be slowly excreted toward outside through the pellicle. Therefore, the extrusomes of O. granulifera might be a kind of excretion extrusomes. Besides, the extrusomes of O. granulifera can be stained by alcian blue 8GS which can rapidly stain mucin, carboxyl and sulphate-containing mucosubstances, therefore, the extrusomes of O. granulifera may contain some mucosubstances. Under the stimulus of methyl green-pyronin, the extrusomes of O. granulifera were numerously discharged, and the extruded contents of extrusomes got together to form a "capsule" which packed the ciliate in it, thus, it might be a defensive response to harmful conditions. Related Articles | Metrics

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Studies on the morphology and histology of head-kidney and spleen in Eleutheronema tetradactylum LUO Jun-biao,XIE Mu-jiao,OU You-jun1,WEN Jiu-fu,LI Jia-er1, WANG Peng-fei 2016, 33 (4):  43.  Abstract ( 345 )   PDF   Save This work focused on histological study on head kidney and spleen in Eleutheronema tetradactylum. The results showed that head kidney, the soft parenchymal organ with brown color and rough surface was located in the backside and around the spinal cord. Under optical microscopy, no renal tubular was found in head kidney encysted by collagen fiber, which filled with numbers of lymphocyte, hematocyte, granulocyte and macrophage, but melano - macrophages center（MC）was not obvious. Spleen shaped as “triangle”, larger in volume and color in brown, staying in junction between intestine and the end of stomach. The spleen was made up of red pulp and white pulp with capsule covering outside, and spleen trabecular structure consisting of expanding connective tissue was obvious. In red pulp, large number of blood cell crowed together being the splenic cord， between this item was splenic sinus, there often had multiple MC; large number of lymphocytes distributed in white pulp with blood vessels going through the center. The features of spleen and head kidney in E.tetradactylum indicated that they are typical immune and hematopoietic organs. Related Articles | Metrics

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Research on the toxicity of glufosinate ammonium to Tetrahymena thermophila DONG Shan-shan, WANG Wei, GENG Ming, WANG Zhan-jun, XU Zhong-dong 2016, 33 (4):  48.  Abstract ( 265 )   PDF   Save The toxicity effects of glufosinate ammonium to Tetrahymena thermophila were investigated from the viewpoints of cell proliferation and DNA damage. The growth curves of 96 hours were plotted by counting the cells every 12 hours; the tail length, tail moment and Olive tail moment of Tetrahymena thermophila were measured by Single Cell Gel Electrophoresis (SCGE). The results of growth curve test showed that there was a significant difference between drug groups and control group in cell density (P＜0.01). According to SCGE data, the tail length, tail moment and Olive tail moment of the tested series were all significantly larger and higher than those in the control group (P＜0.01), the DNA damage level was higher when toxic compound become heavier (P＜0.01). The experimental concentration of glufosinate ammonium caused damages of cell DNA in varied extents, which reflects the acute and genetic toxicity of it to Tetrahymena thermophila . Related Articles | Metrics

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Wetland avian community features of Ziwuling Nature Reserve in Shaanxi XU Shi-cai, SHEN Xue-jian, WANG Li, XI Xiu, BAI Jian-bo 2016, 33 (4):  52.  Abstract ( 239 )   PDF   Save In March 2013 and February 2014, the bird′s number and species of Ziwuling Nature Reserve, Shaanxi was investigated and recorded by line-transect and point count method. And more, the composition, structure, and seasonal dynamics of the avian community were analyzed and researched . A total of 42 bird species belonging to 27 families of 15 orders were recorded. The bird community was mainly composed of Passeriformes, totally 1590 individuals and 18 species, accounting for 47.98% of the total number of bird species. The species(33),number(1245), Shannon-Wiener index (4.94027) and Species evenness(0.97936) in summer were higher than those in the other three seasons, but the Superiority（0.03187） were lowest. Passer montanus, Phasianus colchicus, Pica pica, Cyanopica cyana, Parus major and Upupa epops were the dominant bird species distributed in four seasons, The Sorenson similarity index showed that the lowest species conversion rate was in summer-autumn avian community, the highest was in the autumn-winter avian community. In general, the avian community stability was high in Ziwuling Nature Reserve. Related Articles | Metrics

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Effect of litter size on sustainable energy intake in Eothenomys miletus ZHU Wan-long, YANG Tao, FU Jia-hao, WANG Zheng-kun, CAI Jin-hong 2016, 33 (4):  56.  Abstract ( 336 )   PDF   Save Females in mammals appeared to approach an upper limit to the rate of sustained energy intake/metabolic rate (SusEI/MR) during lactation. However, different species of animals may respond differently to the sustainable limit. Here, the energy metabolism was measured during lactation in female Eothenomys miletus raising litters of natural size (Con), and females whose litter size was manipulated during early lactation to support fewer or more pups (minus pups, MP or plus pups, PP). E. miletus significantly decreased their body mass and increased food intake from early to late lactation， and MP females had lower food intake than the control and PP females. Litter size of the PP group decreased significantly over the period of lactation, and pups were weaned at similar weight to that of the controls. MP females supported a significantly lower litter mass throughout lactation compared with the control and PP females, but during late lactation the pups from the MP group were significantly heavier. Resting metabolic rate (RMR) did not differ significantly between the three groups and the energy intake during peak lactation was 3.56, 3.36 and 3.64 RMR for the control, MP and PP females, respectively. All of the results suggested that female E. miletus reached a plateau in food intake at around 12 g/d during peak lactation, which might signify a limit of SusEI at 3.6×RMR. Related Articles | Metrics

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Pollution features and treatment of simple domestic waste disposal sites in Paihe river basin BAI Yu-fang, WU Ke, WU Dong-biao, CHEN Jun, SONG Zao-ming, MENG Li-li 2016, 33 (4):  60.  Abstract ( 295 )   PDF   Save Through investigation and experimental analysis, the pollution status of the simple domestic waste disposal sites in Paihe river basin was studied, and some measures were put forward. The contents of TP, TN, TOC, CODCr and heavy metals of the leachate respectively were all more than the standard of GB16889-2008. The contents of TP, TN, TOC and CODCr of surface water around simple disposal sites were all more than the standard of GB3838-2002Ⅳ except for TN of Z Township. The contents of heavy metals except Hg in the domestic waste, soil and surface water around simple disposal sites met the standard of GB15618-1995 II or GB3838-2002 IV. The content of Hg in the surface water in which pollution was the most serious was more than about 29 times. The simple domestic waste landfill sites are suitable for local sealing mode to control pollution, on which impermeable membranes are covered and curtain grouting are used. The domestic waste of simple landfill sites, which are far away from sanitary landfill site, are high degree of corruption. The simple dumping site is suitable for the treatment of sanitary landfill in another place, because the domestic waste of simple dumping site, which is located on the farm, is at low level of corruption. Related Articles | Metrics

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Research progress on proteomics of lignocellulose-degrading white-rot fungi WANG Feng-juan, LI Wei-qing, MU Zhi-mei, WANG Yan-wen, LIU Qing-xin, GAO Hui-ju 2016, 33 (4):  65.  Abstract ( 277 )   PDF   Save Lignocellulose is a great potential feedstock for bioenergy production. However the key obstacle for degrading and utilizing lignocellulose, developing biorefinery is the complicated structure of the plant cell wall. White-rot fungi are major degraders of lignocellulose in the nature. Illumination the molecular mechanism of biodegradation would provide a useful foundation for exploring the degradation process of lignocellulose and using the available biomass resource more efficiently. In recent years, some progresses have been achieved in proteomics of lignocellulose-degrading microorganism, providing important information for further exploring the biodegradation mechanism of lignocellulose. This review summarized previous research advance in intracellular proteomics, subproteomics, secretome proteomics and the interactions of degrading enzymes of white-rot fungi in the lignocellulose-degrading process, in order to illuminate its molecular mechanism in degrading lignocellulose from the point of view of proteomics. Related Articles | Metrics

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The research progress of epigenetic regulation of T helper cell differentiation HUAI Wen-ying, YANG Fu-quan, TANG Yu-qin, ZHANG Tian-e, LIU Wei-wei 2016, 33 (4):  70.  Abstract ( 369 )   PDF   Save CD4+ helper T cells (Th cells) differentiate into different cell subsets in order to resist or adapt to the environment changes under the condition of antigen stimulation and body micro-environment, and the process of cells differentiation is controlled by the complex regulatory network of cytokines and transcription factors.Epigenetics is a science that does not involve the change of DNA sequences and changes gene expression such as DNA methylation and chromatin structure, and passes between the parental and filial generation.Epigenetics have an important influence on the differentiation regulation of immune cells and the heritability of T helper cell lineage.  Related Articles | Metrics

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erapeutic strategies of catechins for Alzheimer′s disease aiming at amyloid cascade hypothesis LIU Yan-xia, LAO Feng-xue, YANG Ze-feng, ZHANG Yi 2016, 33 (4):  73.  Abstract ( 334 )   PDF   Save tAlzheimer′s disease (AD), the most common form of dementia among the elderly, is characterized by the progressive decline of cognitive function. Increasing evidence indicates that the production, accumulation and aggregation of amyloid β (Aβ) are central to the pathogenesis of AD. Catechins are widespread in the plant kingdom and have been shown to exert neuroprotective activities in AD involving multimodal activities. The therapeutic strategies of catechins for AD targeting amyloid cascade hypothesis are reviewed and the main problems in the further research are discussed, aiming to providing some references for the study and application of catechins in AD. Related Articles | Metrics

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The molecular mechanism of plant reproductive development YIN Wei-ping, LIU Chuang, PAN Mei-xin, DI Gui-qiu, SHEN Zhe, LIU Xue-mei 2016, 33 (4):  78.  Abstract ( 411 )   PDF   Save The plant life cycle includes two parts: vegetative growth and reproductive growth. The reproductive development can be divided into three stages: the flowering development, the development and formation of gametophyte, fertilization and the formation of zygote. From the survival in the environment and the process of overall development, the development of seed also was part of the plant′s development. Plant growth at different stages has different genes regulatory networks and molecular regulation mechanism, the related gene regulation mechanism was reviewed from the perspective of reproductive growth stages. Related Articles | Metrics

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Effect of tea polyphenols on reproductive performances and its mechanism in sows WU Xiao-min, FENG Tao, BAI Jia-hua, XU Xiao-ling,HUANG Zheng, HAN Xiang-min, LIU Yan 2016, 33 (4):  83.  Abstract ( 374 )   PDF   Save Tea polyphenols is the generic term of polyphenols compounds in green tea, which is a green, new, efficient antioxidants. Tea polyphenols has been applied widely due to its special chemical structure which has unique biological effectiveness. In recent years, it has been found to also play an important role on healthy and efficient breeding. This article reviewed the effect of tea polyphenols on reproductive performance of sows and its mechanism. Related Articles | Metrics

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Advances in the artificially reproductive and breeding studies of silver pomfret, Pampus argenteus HU Jia-bao, YUE Qi-jun, DU Li-hong, ZHENG Hua-kun,XU Xiao-shuang, XU Shan-liang, WANG Ya-jun 2016, 33 (4):  87.  Abstract ( 506 )   PDF   Save As an important marine commercial fish, with the change of environment and human exploitation, the number of wild silver pomfret (Pampus argenteus) increasingly reduces. On the basis of biological characteristics of silver pomfret, the artificial cultivation and breeding will be the main reducing reason. The reproductive periods are from April to June each year, and the spawning areas are mainly in depth of 10-20 m and 26‰-31‰ salinity, close to the river estuary and the coast with abundant nutriment and plankton;the gonadal change of silver pomfret can be divided into phase I ~ VI, and the fish will be different developing period in same time because of temperature and illumination. At present, there is large difference between wild and reared silver pomfret in the development of gonadal tissue, and the ordinary feed is unable to provide adequate nutrition in period of reproduction, so the feed should be added with some supplement such as the copepods and jellyfish; bubbles, white spot and bacterial disease could causes high mortality rate of the fish, hence, in the process of culturing, the cure should be combined with prevention. In this paper, based on current internal and external studies about wild and artificial breeding, we compare the characteristics of gonadal and embryonic development of them, and summarize the research results about culturing parental fish, control of reared environment, component of feed. Based on the current problems existing in the artificial rearing, we put forward some suggestions and the prospect about promotion of culturing quality, nutrition of feed and the screen of qualified strain. Related Articles | Metrics

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The protection of terrestrial wildlife: habitat destruction and mechanism YU Li-li,WANG Hao,SHEN Wen-xing 2016, 33 (4):  92.  Abstract ( 268 )   PDF   Save  The protection of terrestrial wildlife is important for biodiversity protection. Based on the analysis of the relationship between terrestrial wildlife resources and biodiversity, the authors, from the angle of wetland, forest and grassland habitat, analyze the causes of habitat destruction. The results show that production of agriculture, forestry and animal husbandry, industrialization process and excessive consumption are the main reasons.To improve the terrestrial wildlife habitat, the authors,combining with the actual situation,put forwad measures from legal protection,habitat management and guiding rational consumption.  Related Articles | Metrics

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A review of the flow analysis methods of metabolic network PANG Jian, LIU Zhan-ying, HAO Min 2016, 33 (4):  97.  Abstract ( 770 )   PDF   Save The microbial metabolic network is one of important fields of systems biology. The metabolic network flow analysis method has great significance in studying metabolic network properties. Metabolic network flow analysis method is a key to understand the relationship between metabolic network structure and function, metabolic flow and gene expression. It has an important realistic significance in analyzing disease, identify target for drugs, pharmaceutical engineering and genetic modification. Several kinds of metabolic network flow analysis methods were reviewed in the paper in order to supply a reference for analyzing metabolic flux and thus guiding the experiments researches as well. They are flux balance analysis(FBA), minimization of metabolic adjustment(MOMA), regulatory on off minimization(ROOM), dynamic flux balance analysis with linear quadratic regulator(DFBA-LQR), the analysis of flow range(FVA), a bi-level programming framework for identifying gene knockout strategies(OptKnoock), up/down regulations of gene expression(OptReg) and the framework for redesign of production systems(OptStrain).  Related Articles | Metrics

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Optimization of the method for measurement of the activity of small GTP binding protein RhoA ZENG Ting, CUI Zhao-meng, JIANG Wei, LI Zeng-xia, DANG Yong-jun, YU Long 2016, 33 (4):  103.  Abstract ( 1324 )   PDF   Save The small G protein of Rho GTPase family was key regulator in cell migration. As the important member of Rho GTPase family, RhoA regulates the aggregation and contraction of actin, which plays a vital role for cell migration. Monitoring the active RhoA is essential to study RhoA′s function in vivo and as a potential target for tumor therapy. RhoA binding domain (RBD) of Rhotekin, a RhoA effector protein interacts only with active RhoA. RBD pulldown assay is a common assay and usually used to detect GTP-RhoA in vivo. However, gene sequence of RBD with original codon ectopically expressed in E.coli has a relative low expression level which influence the following measurement of active RhoA efficiently. We totally synthesized the sequence of RBD through optimization of mammalian codons to prokaryotic codons, and then cloned RBD into pGEX prokaryotic expression vector. The GST tagged RBD protein was highly expressed in E.coli, and specifically bound to the active form of RhoA (GTP-RhoA). The GST-RBD construct and the optimized pulldown experiment system would provide practical technique for the role of RhoA in cell migration.  Related Articles | Metrics

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The purification of placenta hemoglobin CHEN Chen, WANG Pei, QI Dong-lai, WANG Xiang 2016, 33 (4):  107.  Abstract ( 332 )   PDF   Save The aim of the study is to establish the purification methods of placenta hemoglobin. The placenta hemoglobin was eluted by 0.9% Na chloride(NS) and extracted by distilled water, which was further purified by Q300 anion-exchange chromatography, and then the final products was got by concentrating the purified hemoglobin. The concentration of hemoglobin and the content of methemoglobin of the final products was measured, and the purity was analyzed by SDS-PAGE and HPLC,the oxygen saturation curve was analyzed through the blood analyzer.The concentration of final products was 11.5 g/dl,and the content of methemoglobin is 1% or less; hemoglobin had a higher purity by the analysis of SDS-PAGE and HPLC, excluding mixed protein, the purity was 99.10%±0.70% analyzed by Gel Imaging System; the oxygenation of hemoglobin curve had no significant deviation after purification, the activity of carrying oxygen would not be affected. The research shows that the methods to purify placenta hemoglobin were established. Related Articles | Metrics

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Synthesis of laurate starch through lipase-catalyzed reaction in \[BMIM\]BF4 ionic liquid YUAN Jiu-gang, XIANG Zhong-lin, FAN Xue-rong, WANG Qiang 2016, 33 (4):  110.  Abstract ( 273 )   PDF   Save In order to improve the hydrophobilicity of raw starch, lauric acid was grafted on starch through CALB lipase-catalyzed reaction in \[BMIM\]BF4 ionic liquid. Firstly, the structure of modified starch was tested with 1H NMR, it showed that the lauric acid was successfully grafted on starch. Secondly, the effect of different reaction conditions on the DS values of modified starch was studied in detail. Results showed that a better synthesis condition for laurate starch was as follows: lipase dosage 0.2 g, lauric acid/ AGU=3:1, temperature 60℃，time 4 h. Finally, the hydrophobilicity of modified starch was tested through water contact angle. Results showed that the hydrophobicity of modified starch was significantly improved and increased with DS values.  Related Articles | Metrics

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Reform and innovation on experimental teaching method of veterinary clinical diagnostics WANG Xi-chun, LI Yu, FENG Shi-bin, LI Jin-chun, WU Jin-jie 2016, 33 (4):  114.  Abstract ( 309 )   PDF   Save veterinary clinical diagnosticsis a compulsory basic course of the veterinary profession, which not only contains abundant theoretical knowledge, but also involves a number of experimental skills. It plays a very important role in the talent training of veterinary profession. This article aimed at the problems and shortcomings of traditional experimental teaching, and exploring new teaching ideas which suitable for veterinary professional development through the reforms of platform construction, teaching programs, diagnostic techniques and examination mechanism. The reforms improved the enthusiasm and initiative of students, which achieved good experimental results, and also would provide a reference for reforming of other experimental teaching methods. Related Articles | Metrics

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The general formula about genetic equilibrium of sex-linked gene and its application in teaching DING Jian-Hua， ZHANG Hai-jun 2016, 33 (4):  118.  Abstract ( 435 )   PDF   Save Hardy-Weinberg is one of the theoretical foundations of population genetics, which is not only applicable to the gene balance problem of the autosome, but also applicable to the sex-linked genes. The equilibrium problem of a pair of genes on the autosome is relatively simple and easy to be understood because it is not related to gender, whereas the sex-linked genes do not. The genetic equilibrium characteristics of sex-linked genes were described in detail, and two general formulas were given at the same time in this paper. In the process of teaching, the genetic equilibrium oscillation process of sex-linked genes can be vividly displayed by using the two formulas, and the formula input and mapping function of Excel software, so as to achieve a good teaching effect.  Related Articles | Metrics

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Construction and practice on the innovative practice teaching system of university botany ZHANG Xiu-ling, FU Ai-yun, LI Yan, WANG Li-yan, DAI Zhong-min 2016, 33 (4):  122.  Abstract ( 223 )   PDF   Save Practice teaching is an important portion of botanical teaching. In this paper, according to the training objectives of the biological sciences, aiming at cultivating iwnovative talents， we set up a practical teaching system of botany practice, optimized the practice teaching content, practiced the teaching methods and means，improved the basic teaching facilities and conditions, and implemended the laboratory open operating mechanism. The consciousness capabilities of students are cultivated. Based on the practices, the design of botany practice teaching is discussed in order to provide useful references for the experimental education reform. Related Articles | Metrics

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Reform and establishment of the experiment evaluation system of medical microbiology YANG Cheng， LIU Hui 2016, 33 (4):  125.  Abstract ( 245 )   PDF   Save Medical microbiology experiment， as an important part of medical medical microbiology， focuses on training students′ practical ability and the comprehensive ability to solve the problem. Linking clinical, a new mode of microbiology experiment was reformed. Its goal was to improve students′ comprehensive experimental ability,which the experiment evaluation system was establised to evaluate the effect of the reform.The results showed that the reform can improve the students′ operating skills, cultivate the students′ scientific thinking. The evaluation system can reflect students′ experimental and comprehensive abilities.  Related Articles | Metrics

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Application of internet-based teaching model in teaching of medical biochemistry LUO Yue-ji, ZENG Jie, LI Yan, ZHU Chuang-bing 2016, 33 (4):  127.  Abstract ( 249 )   PDF   Save The internet-based teaching model is the teach-orientated model which improves students′ interest in studying, changes the studying pattern into an active style and strengthens the students′ comprehensive qualities. We change the traditional teaching model into an internet-based style on the basis of the construction of selected courseware of Hunan Province. In the internet-based teaching model, the teaching methods include internet courseware, mock course, the problem-based learning and interactive communication, and the teaching results of which will be analyzed by the final examination and the questionnaire survey. The results states that the internet-based teaching model can improve students′ performance in studying, motivate students′ interest, improve the students′ understanding of course material, learning and innovation skills. In order to get better performance in the future, the internet-based teaching model can be modified and applied in the further teaching activities.  Related Articles | Metrics