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Table of Content

    • Current Biotechnology
      Volume 10 Issue 3 25 May 2020
    • Advances in the Study of Inflammation-tumor Transformation in Ocular Diseases
    • LIU Shibo1, WU Hao1, HONG Jiao2, LIU Mengyu1, YAO Mawulikplimi Adzavon1, ZHAO Pengxiang1*
    • 2020, 10(3):  234-241.  DOI: 10.19586/j.2095-2341.2019.0110
    • Abstract ( ) PDF (1313KB)( )
      • The mechanism of inflammation transforming into cancer has always been the focus of cancer research. As the environment in which inflammation-tumor transformation starts, inflammatory microenvironment is a large collection of various regulatory factors and cells, including tumor stem cells, tumor-related macrophages and cytokines (such as chemokines and growth factors) that play crucial roles in the process of tumor initiation, occurrence, evolution and even malignant transformation and metastasis in common ocular tumors. Based on this, the effects of tumor-related cells, cytokines and extracellular matrix in inflammatory microenvironment on the proliferation, metastasis, infiltration and invasion of tumor cells were mainly discussed, with emphasis on the molecular mechanism related to ocular inflammation-tumor transformation. The characteristics of common ocular tumors, such as retinoblastoma and adenoid cystic carcinoma, and the molecules that play  important regulatory roles in the process from inflammation to tumorigenesis were also reviewed. Finally, the future diagnosis and treatment methods were prospected for the ubiquitous signal pathways and molecular targets of these ocular tumors, so as to design ideas for the mentioned inflammatory components in the future diagnosis and treatment of ophthalmic tumors, and to maximize the prevention of inflammation-tumor transformation and malignant transformation.
    • Screening Locus-specific SSR Markers Associated with the QTL for Scab Resistance in Wheat and Evaluating Their Allelic Variation and Genetic Effects
    • WU Di1,2,ZHENG Tong1, LI Lei1, LI Tao1*
    • 2020, 10(3):  242-250.  DOI: 10.19586/j.2095-2341.2020.0042
    • Abstract ( ) PDF (4306KB)( )
      • Fusarium head blight (FHB) is one of major diseases in wheat. Stacking different quantitative trait loci (QTL) via marker-assisted selection is an effective and environmentally friendly approach to control the disease and to reduce the toxin content in grains. The QTL for scab resistance have been identified throughout the wheat genome, however, except for a few QTL, such as Fhb1 and Fhb2, most of the QTL lack efficient diagnostic markers. simple sequence repeat (SSR) marker is rich in polymorphism, which can differentiate different alleles in a natural population, and can be easily used in marker assisted breeding. Based on this, a panel of 386 SSR markers associated with scab resistance were collected from published literatures, which were then used to construct consistent genomewide QTL maps for scab resistance. Variation of copy numbers for these associated markers were analyzed, and SSR markers of single copy were selected. These single-copy markers were then used to genotype a natural population consisting of 156 varieties, and association of single marker genotypes with phenotypic data collected in field and greenhouse environments for three seasons each were performed. The allelic variations and their genetic effects of those single-copy SSR markers significantly associated with scab resistance were analyzed. The results showed that a total of eight single-copy SSR markers were significantly correlated with the phenotype in at least two seasons (P<0.05), involving chromosomes 2D, 2B, 3B, 5A, 5B, 6A, 6D and 7A, and five single-copy markers had effective alleles. Chinese wheat landraces and Japanese varieties had more favorable (resistant) alleles, and the more favorable alleles the varieties had, the better the resistance to scab was observed. These QTL loci and their linked single-copy SSR markers could be used in wheat breeding to improve the level of scab resistance and breeding efficiency.
    • Alleviating Effect of Tieguanyin Tea Aqueous Extract on Hyperuricemia in Mice
    • XI Yuanyuan§, XU Hongyan§, LIU Xia, YIN Huiqing, LIU Xiaopan, WANG Qinglu, WANG Qinglu*
    • 2020, 10(3):  256-264.  DOI: 10.19586/j.2095-2341.2019.0084
    • Abstract ( ) PDF (7509KB)( )
      • In recent years, hyperuricemia (HUA) occurs frequently in the population, with strong harmfulness and many complications. In order to explore whether semi-fermented  Tieguanyin tea, a common daily drink, has an auxiliary effect on alleviating HUA, mice were taken as experimental objects, and mice hyperuricemia models were established by the combined method of potassium oxazinate and hypoxanthine. Twenty-one model mice were randomly divided into model group, Tieguanyin tea aqueous extract group, positive drug group, and 7 non-model mice as controls. After 2 weeks of different treatments, mice serum, kidney, liver and small intestine were taken. The pathological changes of various organs were observed, so as to identify the effects of Tieguanyin tea aqueous extract on various organs of hyperuricemia mice from the cellular level. Biochemical indicators with high correlation with HUA were measured: concentration of serum uric acid (UA) and activity of xanthine oxidase (XOD) in liver to determine whether the model was successfully established and whether Tieguanyin tea aqueous extract had a mitigating effect on HUA. qRT-PCR was used to detect the mRNA expression level of genes related to uric acid synthesis and excretion, and Western blot was used to detect the expression level of liver XOD protein, so as to clarify the influence of Tieguanyin tea aqueous extract on HUA from the molecular level. Pathological sections showed that compared with the positive drug group, the mice in Tieguanyin tea aqueous extract group had less kidney and liver damage. Biochemical indexes showed that Tieguanyin tea aqueous extract could reduce serum uric acid level and inhibit XOD activity. From the molecular level, it could be seen that Tieguanyin tea aqueous extract significantly increased the mRNA expression levels of uric acid transporter 1 (URAT1) and organic anion transporter 3 (OAT3) (P<0.05), and the expression of glucose transporter 9 (GLUT9) and organic anion transporter 1 (OAT1) was significantly reduced (P<0.05). Although qRT-PCR and Western blot indicated that the expression level of XOD mRNA and protein increased, but the production of uric acid decreased, suggesting that a certain component in Tieguanyin tea aqueous extract may increase the expression of XOD protein without catalytic activity, thus causing a positive feedback to the transcription and expression of XOD gene. The research indicated that Tieguanyin tea aqueous extract had an alleviating effect on hyperuricemia in mice, and its alleviating effect was related to inhibiting XOD activity, intervening uric acid production process and stimulating or inhibiting mRNA expression of related anion transporters.
    • Data Mining Analysis of Prognosis Related Genes of Glioblastoma
    • MA Shengnan, ZHANG Xiaokang, YI Yang, ZHANG Zhao, YAO Tingting, ZHAO Qinghui, XIE Fei*
    • 2020, 10(3):  265-272.  DOI: 10.19586/j.2095-2341.2020.0025
    • Abstract ( ) PDF (11605KB)( )
      • Glioblastoma (GBM) is the most aggressive human brain tumor. Since currently available drugs are of limited efficacy, the recurrence rate is high and the prognosis is poor for GBM. There is an urgent need to develop a new anti-GBM drugs in clinical practice. The purpose of this study was to screen genes that are closely related to the prognosis of GBM and to provide clues for finding new drug targets. In the study, the GEO2R tool was used to initially screen differentially expressed genes from 269 tumor tissues and 61 normal tissues in the GEO database. Cluster Profiler databases was then used to perform the gene function enrichment analysis, STRING and Cytoscape further resulted a total of 37 differentially expressed genes. The expression of these genes were then verified by GEPIA interaction analysis in GBM tumor tissues. To further explore the relationship between these differentially expressed genes and the prognosis of patients with GBM, GEPIA was used to deeply mine the patient prognosis-related data in the TCGA database. Finally, three genes, named PTTG1, RRM2, and E2F7, were found to be significantly negatively correlated with the patient's median survival. Conclusively, the screened genes highly related to patient prognosis not only offer a reference for assessing patient prognosis, but also provide potential targets for the development of new anti-GBM drugs.
    • Expression and Function of KIF26B in the Development of Non-small Cell Lung Cancer
    • LI Fangfang, MU Dengcai,YANG Chunyan, WANG Lei, SHEN Hao, ZHENG Shangyong*
    • 2020, 10(3):  273-283.  DOI: 10.19586/j.2095-2341.2019.0132
    • Abstract ( ) PDF (13763KB)( )
      • Although kinesin is related to tumorigenesis closely, whether it is related to the expression and development of non-small lung cancer, and its specific biological effects and the molecular mechanisms have been few reported yet. After interfering with KIF26B, we explored the effects on proliferation, invasion, migration, cell cycle, apoptosis, and expression of related proteins in non-small cell lung cancer. By analyzing the mRNA TCGA database information, it was concluded that KIF26B gene is highly expressed in non-small cell lung cancer. Then the expression level of KIF26B in several common non-small cell lung cancer cell lines was detected by qRT-PCR, and KIF26B was screened for high expression in A549 and NCI-H292 cell lines. After using RNA interference (siRNA) to silence the KIF26B gene of A549 and NCI-H292 cells, the silenced KIF26B gene was detected by CCK8, real time cell analyzer (RTCA), plate cloning and Transwell experiments. Flow cytometry was used to analyze the effects of silenced KIF26B gene on cell cycle and apoptosis in NCI-H292 and A549, and Western blotting was used to detect the protein expression. The results showed that after silenced KIF26B, the proliferation of A549 and NCI-H292 cells was significantly reduced, and the ability to invade and migrate was significantly reduced. After silencing KIF26B, A549 and NCI-H292 cells were prevented from transforming from G1 to S phase, and apoptotic cells were significantly increased. Besides, the protein expression level of related cyclin D1, Bcl-2, E-cadherin and Vimentin reduced significantly, while activated Caspase-3 (active Caspase-3) and its cleavage substrate PARP1 (cleaved PARP1) expression levels were significantly up-regulated. The results indicated that KIF26B may be a tumor-promoting gene for non-small cell lung cancer, and it is involved in the occurrence and development of non-small cell lung cancer, KIF26B is expected  to be a potential target for the treatment of NSCLC.
    • Application of Antibody Titration in Flow Cytometry of Hematopoietic Cells
    • ZHANG Xiaoru1, ZHANG Mingying1, LIU Yining1, FU Lixia2, XING Wen1, ZHOU Yuan1*
    • 2020, 10(3):  284-291.  DOI: 10.19586/j.2095-2341.2020.0014
    • Abstract ( ) PDF (6586KB)( )
      • Flow cytometry is one of the important methods to study and diagnose hematological diseases, which can realize high-speed quantitative analysis and sorting distinct cell populations. However, due to different cells and different experimental conditions used in different experiments, non-specific staining of antigen-negative cells often occurs. Using antibody titration method, the experimental conditions for the optimal separation of antigen positive cell population and negative cell population can be obtained by calculating and comparing staining indexes. In order to optimize the experimental conditions for staining with fluorescent dye conjugated antibodies in flow cytometry, mouse bone marrow cells were taken as labeled cells, and Rat Anti-Mouse CD11b antibody labeled with non-tandem fluorescent dye FITC (FITC Rat Anti-Mouse CD11b) and Rat Anti-Mouse CD11b antibody labeled with tandem fluorescent dye APC-efluor 780 (APC-eFluor780 Rat Anti-Mouse CD11b) were selected for labeling. By calculating the staining index of mouse bone marrow cells labeled with serially diluted antibodies, antibody titration was carried out to determine the appropriate antibody concentration interval, and then the effects of cell number, staining time and fixing steps on antibody staining index were analyzed to explore the key factors affecting antibody staining of blood cells. The results showed that the staining index of FITC Rat Anti-Mouse CD11b and APC-eFluor780 Rat Anti-Mouse CD11b were higher in the range of 0.156~2.500 μg·mL-1 and 0.25~1.00 μg·mL-1, respectively, but the antibody concentration beyond this range would reduce the staining index. When the antibody concentration and staining time were constant, the FITC Rat Anti-Mouse CD11b and APC-eFluor780 Rat Anti-Mouse CD11b have higher staining index in the range of 1.56×105~5.00×106 cells·tube-1 and 1.56×105~3.12×105 cells·tube-1 respectively, but the number of cells beyond this range would reduce the staining index. When the concentration of antibody and the number of cells were constant, the staining index of FITC Rat Anti-Mouse CD11b decreased with the extension of staining time, while APC-eFluor780 Rat Anti-Mouse CD11b was opposite. By comparing the staining indexes before and after fixation, it was found that the staining indexes of FITC Rat Anti-Mouse CD11b and APC-eFluor780 Rat Anti-Mouse CD11b both decreased significantly after fixation (P<0.01 and P<0.05). The study provided a method for optimizing flow analysis of blood cells through antibody titration, and pointed out that selecting appropriate antibody concentration, cell number, staining time and fixing steps according to antibody titration results in specific experiments was of great importance to the study of flow detection of labeled blood cells.
    • Study on the Electrochemical Behavior of Dopamine at Carboxylated Graphene/Cysteamine Modified Gold Electrode
    • GAO Xianjuan1, JI Xia1, WANG Qinglu1, DU Qingqing1, WANG Fengdan1, WANG Huaisheng2
    • 2020, 10(3):  292-298.  DOI: 10.19586/j.2095-2341.2020.0013
    • Abstract ( ) PDF (1813KB)( )
      • In order to establish a more advanced dopamine biosensor based on the carboxylated graphene/cysteamine modified gold electrode for quantitative detection of catecholamine neurotransmitter dopamine, cysteamine was modified on gold electrode by self-assembly technology, and then the carboxylated graphene was fixed on the modified electrode by 1-ethyl-\[3-dimethylaminopropyl\] carbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS) crosslinker to prepare the dopamine electrochemical sensor. Firstly, the modified electrode was characterized to test its sensitivity, and then the electrochemical behavior of the electrode in dopamine solution was studied by cyclic voltammetry, including optimization of detection conditions and determination of sensor performance. Characterization results of the modified electrode showed that the carboxylated graphene/cysteamine modified gold electrode improved the electron transfer ability of the electrode and had higher sensitivity. Through single factor experiments, the best detection condition was to use 0.30 mol·L-1 phosphate buffer solution (PBS) with pH 6.00 to prepare dopamine solution, and the scanning rate  set as 200 mV·s-1. Under the optimal detection conditions, the magnitude of the current of the prepared dopamine electrochemical sensor increased with the increase of dopamine concentration, showing a good linear relationship in the range of 1.0×10-3 ~ 3.5×10-3 mol·L-1. The linear regression equation was I=8.120 6C+7.017, and the correlation coefficient R2 was 0.999 5. Moreover, the sensor had good precision, strong stability and certain anti-interference capability. The research results provided support for the determination of dopamine content in drug dopamine hydrochloride injection.
    • Microalgae Breeding with High Efficient of Oil-producing and Optimization of Cultivation Conditions
    • ZHANG Hongbing, LIU Hui, SHI Xiuying, LI Huixuan, FAN Daochun
    • 2020, 10(3):  311-319.  DOI: 10.19586/j.2095-2341.2020.0011
    • Abstract ( ) PDF (6397KB)( )

      • It is necessary to screen microalgae strains that can adapt to the climate of Shijiazhuang because the feasibility of using microalgae in wastewater treatment as a supplement for wastewater treatment can couple between environmental pollution control and the alleviation of fossil energy shortage. In this paper, 10 microalgae strains were selected after purification from rivers in Shijiazhuang city by using BG11 medium. The data of all microalgae strains culture were shown that the DK-3 was the best one than others with the high biomass(1.86 g·L-1) and the high total fat content \[(10.23±2.63) mg·L-1·d-1\] under the autotrophic condition by variance analysis with SPSS 19.0 and LSD multiple comparison. The DK-3 was identified as Scenedesmus meyen and the phylogenetic tree was constructed by 18S rDNA sequencing analysis. In order to explore the optimal heterotrophic conditions of algae DK-3, the experiment scheme was designed by couple between Box-Behnken design (BBD) and response surface method (RSM). The result showed that the best culture condition of DK-3 was the BG11 with glucose 20 g·L-1, K2HPO4·3H2O 90 mg·L-1, and MgSO4·7H2O 200 mg·L-1, respectively. Compared with the biomass (3.37 g·L-1) of unoptimized group, the biomass (5.47 g·L-1) and oil yield (1.28 g·L-1) of DK-3 under the optimized conditions above was increased by 62.31% after 240 h training. Therefore, the DK-3 is a good algae with a high research potential.

    • Study on Fermentation Process of Lycopene Production
    • WANG Yingyan, CHEN Wenna*, JIA Cunjiang, FANG Shubiao, QIN Bin, ZHU Zhengqiao
    • 2020, 10(3):  320-327.  DOI: 10.19586/j.2095-2341.2019.0124
    • Abstract ( ) PDF (7032KB)( )
      • Lycopene is a kind of carotene, which exists in many fruits and vegetables. It has good antioxidant, anti-aging and immunity enhancing functions. It is widely used in medicine, food, cosmetics and other fields. However, the traditional extraction method of lycopene has the disadvantages of low yield and high cost, and it is limited by the source of raw materials. Chemical synthesis method has potential safety hazards. Microbial fermentation has attracted increasing attention due to its advantages of simple process, high production efficiency, and the same biological activity as natural plant extracts. Based on this, taking lycopene yield as response value, response surface methodology was used to optimize metabolic control conditions on the basis of single factor experiments, and the effects of positive and negative strain inoculation ratio, metabolic flow inhibitor and exogenous vegetable oil on lycopene yield were investigated. The results showed that the optimal conditions were as follows: the inoculation ratio of the negative and positive strain of Braxella trisporus was 6∶1, the addition of lycopene cyclase inhibitor 2,6-dimethylpyridine was 0.09 g·L-1, and 3.57 g·L-1 soybean oil was used as exogenous vegetable oil. Under the optimal conditions, the lycopene yield in the fermentation broth reached 1.51 g·L-1, and the bacterial biomass was 51 g·L-1. The research significantly increased the biomass of fermented bacteria, promoted the conversion rate of lycopene biosynthesis, and provided a theoretical basis for the industrial production of lycopene.

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