Current Biotechnology

Recent Advances of G-protein Research in Maize
WANG Yali1,2, DENG Dexiang1,2, WANG Yijun1,2*
2019, 9(1): 1-5. DOI: 10.19586/j.2095-2341.2018.0093
Abstract ( ) PDF (1130KB)( )

G-proteins comprise Gα, Gβ and Gγ subunits. Gγ subunits could be further classified into three types: Type A, Type B and Type C. Plant G-proteins are involved in various biological processes, including plant architecture determinant, biotic and abiotic stress response, nutrient-use efficiency, and yield performance. Currently, G-protein research in maize is limited in canonical Gα and Type C Gγ subunits. Here, we summarized recent advances of G-protein research in maize with focused on the introduction of agronomic relevance for G-proteins. We also proposed future research in maize G-protein field, which was expected to provide reference for G-protein research in maize.

Progress on Models of Traumatic Brain Injury
ZHAO Qinghui, JU Fangdi, SU Zehua, LUO Qiuli, YI Yang, ZHANG Xiaokang, ZHANG Xin, XIE Fei*, MA Xuemei*
2019, 9(1): 6-12. DOI: 10.19586/j.2095-2341.2018.0067
Abstract ( ) PDF (1099KB)( )

Traumatic brain injury (TBI) not only has higher morbidity and mortality, but also leads to varying degrees of impairment in cognitive activities and sensorimotor functions of its survivors. Establishing a reasonable TBI model helps to understand the pathophysiology of TBI and explore its treatment strategies. Many animal models of traumatic brain injury (belong to in vivo TBI models) have already been used to replicate various traumatic brain injuries in human. Unfortunately, most of the therapeutic regimens with neuroprotective effects in animal experiments proved to be ineffective in clinical studies. Since cells cultured in vitro are not adulterated with complex factors which exit in vivo, in vitro models of various traumatic brain injuries have been set up progressively. According to different types of injury, the common-used in vivo animal models and in vitro cell models are divided into physical injury model, pressure injury model, blast-induced injury model and repeated mild injury model. The characteristics of the above common-used TBI models were summarized and comparatively analyzed in order to help for the search of clinical strategies with neuroprotective effects.

Progress on the Antitumor Activity of Baicalin and its Mechanism
WANG Tingting, ZHAO Pengxiang, XIE Fei *
2019, 9(1): 13-20. DOI: 10.19586/j.2095-2341.2018.0096
Abstract ( ) PDF (2938KB)( )

Scutellaria baicalensis Georgi is one of the most important Chinese herbal medicines in China. It has a large production and a wide range of uses. Baicalin is one of the most important active ingredients and belongs to flavonoids. In traditional Chinese medicine, baicalin has the functions of clearing away heat, detoxifying, reducing blood pressure and antivirus. In recent years, it has been found that baicalin also plays an active role in antitumor. This article focused on the role of baicalin in the antitumor aspects, which was expected to expand the understanding of the efficacy of baicalin, and provide new ideas for the future treatment of cancer.

Function of Glucose Transporter in the Regulation of Blood Glucose Homeostasis
ZHANG Kaiyi1, XIE Ning1, YE Huaqiong1,2, YANG Shulin1*
2019, 9(1): 21-27. DOI: 10.19586/j.2095-2341.2018.0101
Abstract ( ) PDF (1753KB)( )

Glucose transport depends on glucose transporter (GluT), which is the major step of tissue glucose metabolism and blood glucose homeostasis. Class I of GluT family is closely related to metabolic diseases such as T2DM. This paper reviewed the characteristics and functions of Class I of GluT and how they participate in glucose metabolism, blood glucose homeostasis and pathological process of T2DM. The paper also analyzed the characteristics, division and the relationship of the tissues and organs in glycose metabolism. The paper was expected to show new view on diagnosis and treatment of T2DM and related diseases, furthermore, provide new ideas for exploring potential drug target.

Progress on Purification and Clinical Applications of Human Transferrin
HE Jie, GUO Caiping*
2019, 9(1): 28-34. DOI: 10.19586/j.2095-2341.2018.0081
Abstract ( ) PDF (2454KB)( )

Human transferrin is one of the most abundant proteins in human plasma. Its the major iron transporting protein in human plasma and extracelluar space in tissue. And it transports iron from the place of adsorption and storage to the forming red blood cells, where hemoglobin is synthesized, as well as to other requiring iron positions such as dividing cells. Transferrin is an important antioxidant and anti-microbial protein. Also, it could be applied as an additive in serum-free culture to enhance cell proliferation. Furthermore, transferrin has attracted much attention and shown comprehensive prospects in various clinical applications such as congenital atransferrinemia/hypotransferrinemia, iron binding/regulatory therapy and cancer therapy. Herein, the structure characteristics and physiological function of human transferrin were described. In addition, progress of the purification technique and clinical applications of transferrin were reviewed, which was expected to promote the clinical application of transferrin.

Cloning and Functional Analysis of MYB Transcription Factor AaBPF1 in Artemisia annua L.
MA Jiawei, WANG Yuting, YAN Zhenning, FU Xueqing, ZHAO Jingya*
2019, 9(1): 35-45. DOI: 10.19586/j.2095-2341.2018.0090
Abstract ( ) PDF (8437KB)( )

Artemisinin is an important sesquiterpene lactone endoperoxide which is isolated from Chinese medicinal plants Artemisia annua L.. Artemisinin-based combination therapies (ACTs) are recommended as the best treatment for malaria. However, the content of artemisinin in A. annua is low (0.01%~1% of dry weight). Light is a crucial physical factor for plant growth and development. More importantly, it is a significant regulator in the biosynthesis of secondary metabolites. Several studies show that light could regulate the biosynthesis of artemisinin, but its regulatory mechanism is still unknown. According to analysis of a large set of RNA sequencing (RNA-Seq) data of A. annua with red light (40 μmol/m2·s) treatment, AaBPF1, one MYB transcription factor, was screened. The AaBPF1 gene contained a 1 932 bp open reading frame, which encoded a protein with 643 amino acids, and its expression level was induced by red light. The expression pattern of AaBPF1 was similar to those of synthase genes for artemisinin. Subsequently, the expression of synthase genes for artemisinin was detected by Dual-Luciferase Reporter Assay System, and the expression of the target genes and the content of artemisinin in transgenic A. annua plants were analyzed. The results showed that AaBPF1 could significantly activate the expression of ADS and CYP71AV1. The study demonstrated that the transcription factor AaBPF1 might positively regulate the biosynthesis of artemisinin by activating the expression of synthase genes for artemisinin.

Study on Expression and Functional Analysis of GhCAD6 Gene in Transgenic Cotton
HU Wenran1, LI Xiaodong2, ZHOU Xiaoyun1, LI Xiaorong1, YANG Yang1, LI Bo1
2019, 9(1): 46-53. DOI: 10.19586/j.2095-2341.2018.0112
Abstract ( ) PDF (2256KB)( )

To study the expression and function of GhCAD6 gene in transgenic cotton fibers, the transgenic material was obtained by Agrobacterium-mediated transformation into Xinluzao 36, and the T6 generation plant material was obtained after successive generations of selection. The genomic DNA from cotton leaf of the T6 generation material plants was detected by PCR, and the positive plants were identified. The genomic DNA of the positive plants were hybridized by Southern blotting. The results showed that GhCAD6 was integrated into the cotton genome in different genes loci in the form of single copy. Total RNA from cotton fiber was extracted by hot borate-proteinase K method. The method for detecting the expression of GhCAD6 gene in cotton fiber was established by quantitative real-time PCR. The expression of GhCAD6 gene in cotton fiber at different developmental stages was determined by the method. The expression of GhCAD6 in transgenic plant fiber was lower than that in control samples at the same developmental stage. The expression of GhCAD6 was inconsistent with that in the control samples at 15 DPA (day post-anthesis), and the other periods were approximately the same. The change of GhCAD6 gene expression at 15 DPA may be the main reason for the variation of fiber quality of transgenic cotton. The results of this research could provide theoretical basis for further analysis of the mechanism of GhCAD6 gene in improving cotton fiber quality.

Study on Interaction Between Gallic Acid and Bovine Hemoglobin
SHI Shenjia1,2, LI Jianying1,2, LI Zhongbao1*, CHEN Junde2*, WU Kunyuan2,3
2019, 9(1): 54-61. DOI: 10.19586/j.2095-2341.2018.0103
Abstract ( ) PDF (4492KB)( )

Bovine hemoglobin (BHb) is highly homologous to human hemoglobin and obtained easily, so it is often used in the study of binding of hemoglobin to small molecule compounds. Gallic acid (GA) is a polyphenolic small molecule compound, which can be used as drug model. The study on interaction between GA and BHb can provide a theoretical basis for the application of GA in the field of medicine. The interaction between GA and BHb was investigated at pH 7.0 by ultraviolet-visible (UV-vis) spectroscopy, fluorescence spectroscopy, Fourier transform infrared (FTIR) spectroscopy and circular dichroism (CD). The results of UV-vis spectroscopy showed that GA had no effect on oxygenation state of BHb, and did not deoxidize the oxygenated hemoglobin. Combined fluorescence spectra with calculation results of related formulas, it could be known that the interaction between GA and BHb resulted in fluorescence quenching, and the quenching constant increased with the increase of temperature, indicating that the quenching type of the interaction between GA and BHb was dynamic quenching. Then the binding constants of GA and BHb at different temperatures were calculated, which were K298a=7.941×103 L/mol, K308a=10.478×103 L/mol, respectively. And the binding process was a spontaneous molecular interaction procedure, in which hydrophobic interactions played major roles. The results also demonstrated that the microenvironment of tryptophan and tyrosine residues had been disturbed, under the same condition, variations of fluorescence absorption peak intensity of the tryptophan residue were greater than that of the tyrosine residue, indicating that the binding site of GA and BHb was closer to the tryptophan residue. The results of FTIR spectroscopy and CD showed that the secondary structures before and after the interaction between BHb and GA were both dominated by α-helices, it meant that GA had little influence on the secondary structure of BHb molecules.

Screening, Identification of Catechol Siderophore-producing Bacteria from Human Gastrointestinal Tract
GONG Qiang1, ZHANG Xichun2*
2019, 9(1): 62-68. DOI: 10.19586/j.2095-2341.2018.0053
Abstract ( ) PDF (2024KB)( )

In order to screen and identify the facultative anaerobes producing catechol siderophore from adult feces, the facultative anaerobic microorganisms from human gastrointestinal tract were isolated and purified by artificial simulated gastrointestinal tract environment. The positive strains which contained the genes of nonribosomal peptide synthetase (NRPS) in the biosynthesis pathway of catechol siderophore were screened and identified with 16S rRNA sequence. And then the types of products were predicted by the substrate specificity of the NRPS adenin domains, and the abilities of strains to produce catechol siderophore were determined by Rious method. Finally, 8 strains of catechol siderophore-producing bacteria were isolated, which belonged to Bacillus and Escherichia respectively, and the products were bacillibactin and enterobactin. Among them, Bacillus cereus Gut 16 possessed the strongest ability of producing catechol siderophore (94.75±0.40 μmol/L). The result of this research laid a foundation for the study on the function of the catechol siderophore-producing bacteria in the gastrointestinal nutrition and health.

Isolation and Identification of a Nitrite Bacterium and its Optimization of Fermentation Process
ZHAO Pengnian, YANG Deyu, WANG Jiayou, DING Yifan, AN Peng, WANG Yuan*
2019, 9(1): 69-77. DOI: 10.19586/j.2095-2341.2018.0084
Abstract ( ) PDF (1579KB)( )

Nitrite bacteria (also known as ammonia-oxidizing bacteria) are widely distributed in nature. They exist in soil, freshwater and oceans. And nitrite bacteria play important roles in degradation of ammonia nitrogen in sewage. However, due to the difficulty in high-density fermentation of nitrite bacteria, their practical applications are limited. Based on this situation, marine sludge was collected and enrichment cultured in shake flask, and then a nitrite bacterium was isolated from marine sludge by silicone plate and washed agar plate, which was identified as Nitrosomonas sp.. The optimum culture conditions of the strain of nitrite bacterium was determined by shake-flask culture: The optimum temperature is 30℃，the optimum pH is 8.0, and the best dissolved oxygen ≥1.8 mg/L. And the optimal combination and concentration of the medium were determined by orthogonal test, which was 0.5% NaHCO3, 0.8%(NH4)2SO4, 0.1% KH2PO4, 0.01% FeSO4. At last, under the optimal culture conditions, the high-density fermentation of the nitrite bacterium was performed by using 10 L fermentor, and the fermentation broth was replaced twice during the fermentation. The nitrosation rate of the fermentation broth reached 236 mg(N-NO-2)/L·h within a 75-hour fermentation cycle, and it was 2 times higher than that of fermentation batches without fermentation broth replacement. The research showed that replacement of fermentation broth greatly improved the fermentation efficiency, and played an important role in guiding the subsequent industrial production.

Effect of two Brewing Methods on Quality and Anti-oxidation Activity of Rice Wine Adding Hibiscus sabdariffa L.
GE Tingting, GAO Yu, ZHENG Daheng*, ZHENG Panxing, SUN Qian, MA Runyao, LI Qiuxia, HAN Jiwei*
2019, 9(1): 78-83. DOI: 10.19586/j.2095-2341.2017.0156
Abstract ( ) PDF (1275KB)( )

In this paper, quality and anti-oxidation activity of rice wine adding H. sabdariffa were evaluated to compare different brewing methods. The results showed that contend of alcohol, total sugar, amino acid nitrogen and volatile ester adding H. sabdariffa during after-fermentation was higher than pure rice wine（P<0.05, P<0.01）. Moreover its content of total polyphenols and total flavonoids was higher, the difference was significant （P<0.01）. For the rice wine adding H. sabdariffa during after-fermentation, the scavenging rate of DPPH radical, hydroxyl radical and ABTS+· radical was higher than pure rice wine. Results suggested the quality and anti-oxidation activity of rice wine adding H. sabdariffa during fermentation were better.

Study on Design of Colchicine Inhibitors of Tubulin for the Treatment of Rheumatoid Arthritis Based on Computer Simulation
SUN Jing1, LIU Dan2*, HUI Wenqi2
2019, 9(1): 84-88. DOI: 10.19586/j.2095-2341.2018.0007
Abstract ( ) PDF (4341KB)( )

Joint application of non-steroidal anti-inflammatory drugs and Colchicine has good drug effect on the treatment of acute arthritis. But Colchicine can cause serious side effects, including diarrhea, nausea, cramps, abdominal pain and vomiting. In this study, molecular docking simulation technology was used to design the bioelectron and other row inhibitors derived from Colchicine. All of the inhibitors were designed according to Lippinskis five (18) rules, so they all showed good drug and drug similar scores. Results showed that, the COL derivatives of all designs have a greater affinity than COL, and their physical and chemical properties are improved comparing with the female COL.

Research on the CGIAR’s Contribution to the Global Agriculture Based on Bibliometrics
LU Yao§, HUANG Dandan§, WANG Ting*
2019, 9(1): 89-97. DOI: 10.19586/j.2095-2341.2018.0130
Abstract ( ) PDF (4633KB)( )

Evaluating the contribution of Consultative Group on International Agricultural Research (CGIAR) to the global agricultural development has great guiding significance to China to make decisions about the partners and fields when it conducts cooperative research with CGIAR. We made bibliometrics analysis on the journal articles that CGIAR published between 2008 and 2017, also evaluated the contributions to the international agricultural research made by CGIAR and its affiliated research centers from the perspectives of article quantity and impact, the main contributing countries and institutes and discipline distribution features. At last, the paper proposed the key fields in which China should strengthen cooperation with CGIAR.

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