1.Effect of wheat-grain moxibustion on Wnt/β-catenin signaling pathway in bone marrow cell in mice with bone marrow inhibition.
Tao ZHU ; Yan-Ting CHENG ; Yan-Zhu MA ; Shuai ZHAO ; Xia LI
Chinese Acupuncture & Moxibustion 2023;43(1):67-71
OBJECTIVE:
To observe the effect of wheat-grain moxibustion at "Dazhui" (GV 14), "Zusanli" (ST 36) and "Sanyinjiao" (SP 6) on Wnt/β-catenin signaling pathway in bone marrow cell in mice with bone marrow inhibition, and to explore the possible mechanism of wheat-grain moxibustion in treating bone marrow inhibition.
METHODS:
Forty-five SPF male CD1(ICR) mice were randomly divided into a blank group, a model group and a wheat-grain moxibustion group, 15 mice in each group. The bone marrow inhibition model was established by intraperitoneal injection of 80 mg/kg of cyclophosphamide (CTX). The mice in the wheat-grain moxibustion group were treated with wheat-grain moxibustion at "Dazhui" (GV 14), "Zusanli" (ST 36) and "Sanyinjiao" (SP 6), 3 moxa cones per acupoint, 30 s per moxa cone, once a day, for 7 consecutive days. The white blood cell count (WBC) was measured before modeling, before intervention and 3, 5 d and 7 d into intervention. After intervention, the general situation of mice was observed; the number of nucleated cells in bone marrow was detected; the serum levels of interleukin-3 (IL-3), interleukin-6 (IL-6) and granulocyte macrophage colony stimulating factor (GM-CSF) were measured by ELISA; the protein and mRNA expression of β-catenin, cyclinD1 and C-Myc in bone marrow cells was measured by Western blot and real-time PCR method.
RESULTS:
Compared with the blank group, the mice in the model group showed sluggish reaction, unstable gait, decreased body weight, and the WBC, number of nucleated cells in bone marrow as well as serum levels of IL-3, IL-6, GM-CSF were decreased (P<0.01), and the protein and mRNA expression of β-catenin, cyclinD1 and C-Myc was decreased (P<0.01). Compared with the model group, the mice in the wheat-grain moxibustion group showed better general condition, and WBC, the number of nucleated cells in bone marrow as well as serum levels of IL-3, IL-6, GM-CSF were increased (P<0.01, P<0.05), and the protein and mRNA expression of β-catenin, cyclinD1 and C-Myc was increased (P<0.05).
CONCLUSION
Wheat-grain moxibustion shows therapeutic effect on bone marrow inhibition, and its mechanism may be related to activating Wnt/β-catenin signaling pathway in bone marrow cells, improving bone medullary hematopoiesis microenvironment and promoting bone marrow cell proliferation.
Animals
;
Male
;
Mice
;
beta Catenin/metabolism*
;
Bone Marrow/physiopathology*
;
Bone Marrow Cells/physiology*
;
Granulocyte-Macrophage Colony-Stimulating Factor/metabolism*
;
Interleukin-3/metabolism*
;
Interleukin-6/metabolism*
;
Mice, Inbred ICR
;
Moxibustion/methods*
;
RNA, Messenger/metabolism*
;
Triticum
;
Wnt Signaling Pathway
;
Hematopoiesis
2.Detection, isolation, and characterization of a novel impurity from several folic acid products.
Qirong SHEN ; Quan HE ; Yuanjiang PAN ; Cuirong SUN
Journal of Zhejiang University. Science. B 2022;23(7):597-606
Folic acid belongs to the group of water-soluble B vitamins and naturally exists in multiple forms in a wide variety of foods such as legumes, vegetables, liver, and milk (Iyer and Tomar, 2009; Lyon et al., 2020). It is involved in many biochemical reactions critical for cell division, such as purine and pyrimidine biosynthesis, DNA/RNA biosynthesis, and amino acid metabolism (Iyer and Tomar, 2009). Mammals cannot synthesize folic acid and thus they must acquire it from food. Although folic acid is ubiquitous in foods, folic acid deficiency still often occurs due to various causes such as unhealthy diet (Hildebrand et al., 2021; Iimura et al., 2022), disease-related malabsorption (Arcot and Shrestha, 2005), medication-related depletion (Arcot and Shrestha, 2005), or vitamin B12 deficiency (Fishman et al., 2000). Folic acid deficiency has been associated with several health problems, such as anemia (Carmel, 2005; Bailey and Caudill, 2012), cancer (Duthie, 1999), cardiovascular diseases (Wald et al., 2002), neural tube defects in newborns (van der Put et al., 2001), neuropsychiatric dysfunction (Shea et al., 2002), depression (Falade et al., 2021), inflammatory diseases (Suzuki and Kunisawa, 2015; Jones et al., 2019), and eye diseases (Sijilmassi, 2019). To prevent folic acid deficiency, its daily intake (400 μg/d) has been recommended for adults in the European Union, and its increased intake (600 μg/d) is advised for women before and during pregnancy (FAO/WHO, 2002; IOM, 2004). The New Zealand government mandated the fortification of non-organic wheat flour with folic acid in July 2021, and the UK government mandated the fortification of non-wholemeal wheat flour with folic acid in September 2021 (Haggarty, 2021).
Adult
;
Animals
;
Female
;
Flour
;
Folic Acid/metabolism*
;
Folic Acid Deficiency/prevention & control*
;
Food, Fortified
;
Humans
;
Infant, Newborn
;
Mammals/metabolism*
;
Pregnancy
;
Triticum/metabolism*
3.Effect of wheat-grain moxibustion on the expression of 5-HT and cortisol in the serum, and MR and GR in the hippocampus in rats with hypothyroidism complicated with depression.
Ji-Yu ZHAO ; Jing YAN ; Hong-Yang WANG ; Qing-Qing LIU ; Tian-Sheng ZHANG ; Chong-Yao HAO
Chinese Acupuncture & Moxibustion 2022;42(5):525-532
OBJECTIVE:
To observe the effect of wheat-grain moxibustion on behavior, 5-hydroxytryptamine (5-HT) and cortisol in the serum, mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) in the hippocampus in rats with hypothyroidism complicated with depression, and to explore the possible mechanism of wheat-grain moxibustion on improving depression in rats with hypothyroidism.
METHODS:
A total of 32 SPF SD rats were randomly divided into a blank group, a model group, a medication group and a wheat-grain moxibustion group, 8 rats in each group. Except for the blank group, the rats in the remaining groups were treated with intragastric administration of 0.1% propylthiouracil (PTU) suspension at 1 mL/100 g, once a day for 4 weeks to establish the rat model of hypothyroidism, and whether the rats were accompanied with depression-like behavior determined through behavioristics evaluation. The rats in the medication group were intervened with euthyrox at 0.9 mL/100 g, once a day, for 4 weeks; the rats in the wheat-grain moxibustion group were treated with wheat-grain moxibustion at "Dazhui" (GV 14), "Mingmen" (GV 4), "Shenshu" (BL 23) and "Pishu" (BL 20), 7 cones each acupoint, once a day, six times a week for 4 weeks. After the intervention, the depression status was observed by behavioristics test; the contents of thyroid stimulating hormone (TSH), total thyroxine (TT4), 5-HT and cortisol in the serum were detected by ELISA; the protein expressions of MR and GR in hippocampus were detected by Western blot; the expressions of MR mRNA and GR mRNA in the hippocampus were detected by real-time PCR.
RESULTS:
Before the intervention, compared with the blank group, the scores of open field test (OFT) were decreased and the immobility time of tail suspension test (TST) was prolonged (P<0.05); the serum TSH contents were increased and TT4 contents were decreased (P<0.01) in the other three groups. After the intervention, compared with the model group, the vertical score of OFT was increased and the immobility time of forced swimming test (FST) was prolonged in the medication group (P<0.05), while the scores of three items of OFT were increased (P<0.05, P<0.01), and the immobility time of FST and TST was shortened in the wheat-grain moxibustion group (P<0.01, P<0.05). Compared with the medication group, the immobility time of TST and FST in the wheat-grain moxibustion group was shorter (P<0.05, P<0.01). Compared with the blank group, in the model group, the contents of serum TSH and cortisol were increased (P<0.01, P<0.001), while the contents of serum TT4 and 5-HT were decreased (P<0.01, P<0.001). Compared with the model group, the contents of serum TT4 and 5-HT were increased, while the contents of serum TSH and cortisol were decreased in the medication group and wheat-grain moxibustion group (P<0.01, P<0.05). Compared with the blank group, the protein and mRNA expression of MR, GR in the hippocampus in the model group was decreased (P<0.01, P<0.05, P<0.001); compared with the model group, the protein and mRNA expression of MR in the hippocampus in the medication group were increased (P<0.05), and the protein expression of MR, GR and mRNA expression of MR in the hippocampus in the wheat-grain moxibustion group were increased (P<0.05, P<0.01). Compared with the medication group, the expression of MR mRNA in the wheat-grain moxibustion group was increased (P<0.05).
CONCLUSION
Wheat-grain moxibustion could significantly improve thyroid function and depression in rats with hypothyroidism. Its mechanism may be related to up-regulating the protein and mRNA expression of MR and GR in the hippocampus, and then affecting the expression of serum cortisol and 5-HT.
Acupuncture Points
;
Animals
;
Depression/therapy*
;
Hippocampus/metabolism*
;
Hydrocortisone/metabolism*
;
Hypothyroidism/therapy*
;
Moxibustion
;
RNA, Messenger/metabolism*
;
Rats
;
Rats, Sprague-Dawley
;
Receptors, Glucocorticoid/metabolism*
;
Receptors, Mineralocorticoid/metabolism*
;
Serotonin
;
Thyrotropin/metabolism*
;
Triticum/metabolism*
4.Screening and evaluation of saline-alkali-tolerant and growth-promoting bacteria.
Xue SUN ; Yonghua DONG ; Na WANG ; Wenhui CUI ; Xianyan LIAO ; Li LIU
Chinese Journal of Biotechnology 2020;36(7):1356-1364
Salinity is the most important factor for the growth of crops. It is an effective method to alleviate the toxic effect caused by salt stress using saline-alkali-tolerant and growth-promoting bacteria in agriculture. Seven salt-tolerant bacteria were screened from saline-alkali soil, and the abilities of EPS production, alkalinity reduction and IAA production of the selected strains were investigated. A dominant strain DB01 was evaluated. The abilities of EPS production, alkalinity reduction and IAA production of strain DB01 were 0.21 g/g, 8.7% and 8.97 mg/L, respectively. The isolate was identified as Halomonas aquamarina by partial sequencing analysis of its 16S rRNA genes, and had the ability to inhibit the growth of Fusarium oxysporum f. sp., Alternaria solani, Phytophthora sojae and Rhizoctonia cerealis. It also could promote root length and germination rate of wheat seedlings under salt stress. Halomonas aquamarina can provide theoretical basis for the development of soil microbial resources and the application in saline-alkali soil improvement.
Alkalies
;
metabolism
;
Bacteria
;
drug effects
;
genetics
;
Halomonas
;
genetics
;
Plant Roots
;
microbiology
;
RNA, Ribosomal, 16S
;
genetics
;
Salt Tolerance
;
genetics
;
Seedlings
;
growth & development
;
microbiology
;
Soil
;
chemistry
;
Soil Microbiology
;
Triticum
;
microbiology
5.High titer ethanol production from an atmospheric glycerol autocatalytic organosolv pretreated wheat straw.
Liang WANG ; Jianquan LIU ; Zhe ZHANG ; Feiyang ZHANG ; Junli REN ; Fubao SUN ; Zhenyu ZHANG ; Cancan DING ; Qiaowen LIN
Chinese Journal of Biotechnology 2015;31(10):1468-1483
The expensive production of bioethanol is because it has not yet reached the 'THREE-HIGH' (High-titer, high-conversion and high-productivity) technical levels of starchy ethanol production. To cope with it, it is necessary to implement a high-gravity mash bioethanol production (HMBP), in which sugar hydrolysates are thick and fermentation-inhibitive compounds are negligible. In this work, HMBP from an atmospheric glycerol autocatalytic organosolv pretreated wheat straw was carried out with different fermentation strategies. Under an optimized condition (15% substrate concentration, 10 g/L (NH4)2SO4, 30 FPU/g dry matter, 10% (V/V) inoculum ratio), HMBP was at 31.2 g/L with a shaking simultaneous saccharification and fermentation (SSF) at 37 degrees C for 72 h, and achieved with a conversion of 73% and a productivity of 0.43 g/(L x h). Further by a semi-SFF with pre-hydrolysis time of 24 h, HMBP reached 33.7 g/L, the conversion and productivity of which was 79% and 0.47 g/(L x h), respectively. During the SSF and semi-SSF, more than 90% of the cellulose in both substrates were hydrolyzed into fermentable sugars. Finally, a fed-batch semi-SFF was developed with an initial substrate concentration of 15%, in which dried substrate (= the weight of the initial substrate) was divided into three portions and added into the conical flask once each 8 h during the first 24 h. HMBP achieved at 51.2 g/L for 72 h with a high productivity of 0.71 g/(L x h) while a low cellulose conversion of 62%. Interestingly, the fermentation inhibitive compound was mainly acetic acid, less than 3.0 g/L, and there were no other inhibitors detected, commonly furfural and hydroxymethyl furfural existing in the slurry. The data indicate that the lignocellulosic substrate subjected to the atmospheric glycerol autocatalytic organosolv pretreatment is very applicable for HMBP. The fed-batch semi-SFF is effective and desirable to realize an HMBP.
Biofuels
;
Carbohydrates
;
chemistry
;
Cellulose
;
chemistry
;
Ethanol
;
metabolism
;
Fermentation
;
Furaldehyde
;
chemistry
;
Glycerol
;
chemistry
;
Hydrolysis
;
Triticum
6.Progress and application prospects of glutamine synthase in plants.
Wanjun FENG ; Guofang XING ; Xulong NIU ; Chen DOU ; Yuanhuai HAN
Chinese Journal of Biotechnology 2015;31(9):1301-1312
Nitrogen is one of the most important nutrient elements for plants and a major limiting factor in plant growth and crop productivity. Glutamine synthase (GS) is a key enzyme involved in the nitrogen assimilation and recycling in plants. So far, members of the glutamine synthase gene family have been characterized in many plants such as Arabidopsis, rice, wheat, and maize. Reports show that GS are involved in the growth and development of plants, in particular its role in seed production. However, the outcome has generally been inconsistent, which are probably derived from the transcriptional and post-translational regulation of GS genes. In this review, we outlined studies on GS gene classification, QTL mapping, the relationship between GS genes and plant growth with nitrogen and the distribution characters, the biological functions of GS genes, as well as expression control at different regulation levels. In addition, we summarized the application prospects of glutamine synthetase genes in enhancing plant growth and yield by improving the nitrogen use efficiency. The prospects were presented on the improvement of nitrogen utility efficiency in crops and plant nitrogen status diagnosis on the basis of glutamine synthase gene regulation.
Arabidopsis
;
Genes, Plant
;
Glutamate-Ammonia Ligase
;
genetics
;
Nitrogen
;
metabolism
;
Oryza
;
Plants
;
enzymology
;
genetics
;
Triticum
;
Zea mays
7.Analysis of three wheat cytoplasmic male sterile lines mitochondrial DNA by AFLP.
Qidi ZHU ; Xinbo ZHANG ; M EJAZ ; Gaisheng ZHANG ; Huixue CHE ; Shuping WANG ; Qilu SONG ; Shuling YANG ; Longyu ZHANG
Chinese Journal of Biotechnology 2013;29(5):646-656
Cytoplasmic male sterility is an important way to utilize wheat heterosis. The purpose of thisstudy was to identify cytoplasmic type of three wheat male sterile lines. Amplified fragment length polymorphism (AFLP) marker technique was used to analyze the wheat mitochondrial DNA. We isolated mitochondria by differential centrifugation and density gradient ultracentrifugation. The results show that the extracted mitochondrial DNA was pure. It was suitable for PCR and genetic analysis. We got 4 pairs of specific primers from 64 primers combinations. Primer E1/M7 amplified 3 specific fragments in ms(Kots)-90-110. Primer E4/M2 generated 2 specific fragments in ms(Ven)-90-110. Primer E7/M6 amplified 2 specific fragments in ms(S)-90-110. Primer E6/M4 produced 2 specific fragments in ms(Kots)-90-110. Four specific primers could be used to identify three cytoplasmic types of Aegilops kotschyi, Ae. ventricosa and Triticum spelta. It provided the molecular basis to further study the mechanism of wheat cytoplasmic male sterility.
Amplified Fragment Length Polymorphism Analysis
;
methods
;
Cytoplasm
;
metabolism
;
DNA, Mitochondrial
;
genetics
;
DNA, Plant
;
genetics
;
Gene Expression Profiling
;
Genotype
;
Plant Infertility
;
genetics
;
Triticum
;
genetics
8.Extracellular expression of protein elicitor PeaT1 in Bacillus subtilis to enhance drought tolerance and growth in wheat.
Limei WANG ; Xiufen YANG ; Hongmei ZENG ; Dewen QIU ; Lihua GUO ; Zheng LIU
Chinese Journal of Biotechnology 2011;27(9):1355-1362
PeaT1, a protein elicitor from Alternaria tenuissima can promote plant growth and trigger systemic acquired resistance in plants. In order to expand the application of PeaT1, P43 promoter sequence and nprB signal peptide-encoding sequence were cloned from Bacillus subtilis 168 chromosomal DNA. The two sequences and peaT1 gene were spliced by overlapping extension. This product was cloned into the Escherichia coli-B. subtilis shuttle vector pHY300-PLK and the resultant recombinant expression vector (pHY43N- peaTI) plasmid was transformed into B. subtilis WB800. SDS-PAGE and Western blotting analysis showed that protein elicitor PeaT1 was expressed extracellularly in B. subtilis. This recombinant bacterial strain enhanced drought tolerance and promoted seedling growth in wheat.
Adaptation, Physiological
;
drug effects
;
Alternaria
;
chemistry
;
genetics
;
Bacillus subtilis
;
genetics
;
metabolism
;
Droughts
;
Fungal Proteins
;
biosynthesis
;
genetics
;
Genetic Vectors
;
genetics
;
Recombinant Proteins
;
biosynthesis
;
genetics
;
Triticum
;
growth & development
9.Impact of distillage recycling on the glycolysis key enzymes, stress response metabolites and intracelluler components of the self-flocculating yeast.
Lihan ZI ; Chunming ZHANG ; Jiangang REN ; Wenjie YUAN ; Lijie CHEN
Chinese Journal of Biotechnology 2010;26(7):1019-1024
This research aimed to study the effect of distillage recycling on ethanol fermentation, the key glycolytic enzymes and cell composition of the self-flocculating yeast. With the self-flocculating yeast SPSC01 and medium composed of 220 g/L glucose, 8 g/L yeast extract and 6 g/L peptone, continuous ethanol fermentation was carried out at the dilution rate of 0.04 h(-1) with a 1.5 L tank bioreactor. Fermentation broth was collected every 3 days, and ethanol and other volatile byproducts were removed by distillation, but the stillage with high boiling byproducts was recycled to prepare the medium instead of fresh water. The system was run for 20 days, during which ethanol and biomass concentrations in the effluent decreased continuously, indicating the significant inhibition of the high boiling byproducts accumulated within the system. Thus, the activities of the key enzymes of the glycolytic pathway: hexokinase, 6-phosphofructose kinase, and pyruvate kinase were analyzed, and it was observed that all of them were inhibited. Furthermore, the biosynthesis of the stress response metabolites glycerol and trehalose was investigated, and it was found that glycerol production that can protect yeast cells against osmotic pressure stress was enhanced, but trehalose biosynthesis that can protect yeast cells against ethanol inhibition was not improved, correspondingly. And in the meantime, the biosynthesis of the major intracellular components proteins and hydrocarbons was adjusted, correspondingly.
Bioreactors
;
microbiology
;
Ethanol
;
metabolism
;
Fermentation
;
Flocculation
;
Glycerol
;
metabolism
;
Glycolysis
;
Hexokinase
;
metabolism
;
Industrial Microbiology
;
methods
;
Phosphofructokinase-1
;
metabolism
;
Saccharomyces cerevisiae
;
enzymology
;
genetics
;
metabolism
;
Schizosaccharomyces
;
enzymology
;
genetics
;
metabolism
;
Trehalose
;
metabolism
;
Triticum
;
metabolism
;
Zea mays
;
metabolism
10.Molecular cloning and preliminary analysis of TaNOA in common wheat.
Lifang HAO ; Chunmei YU ; Bin LI ; Daowen WANG
Chinese Journal of Biotechnology 2010;26(1):48-56
Nitric oxide (NO) is an important signaling molecule with diverse physiological functions in both animal and plant cells. In this work, we isolated the full-length cDNA and genomic DNA sequences of TaNOA-B1 encoding a putative NO associated (NOA) factor in common wheat. Bioinformatic analysis showed that TaNOA-B1 possessed a similar intron/exon structure as its orthologous genes in Arabidopsis and rice. The amino acid sequence deduced from TaNOA-B1 was more than 60% identitical to those of Arabidopsis and rice NOA1 proteins. The primary structure of TaNOA-B1 contained the zinc finger and P-loop GTPase motifs conserved in Arabidopsis and rice NOA1 proteins. There existed at least three NOA gene members in common wheat, which were mapped to homoeologous group six chromosomes 6A, 6B and 6D, respectively. TaNOA-B1 investigated in this work was located on chromosome 6B. The transcripts of TaNOA members were found mainly in leaves. TaNOA-B1-GFP fusion protein may be located in mitochondria. TaNOA transcript level was up-regulated by abscisic acid (ABA) or NaC1 treatments, indicating that TaNOA might be involved in wheat responses to abiotic stresses.
Amino Acid Sequence
;
Arabidopsis
;
genetics
;
Cloning, Molecular
;
DNA, Plant
;
genetics
;
Exons
;
Genes, Plant
;
genetics
;
Introns
;
Molecular Sequence Data
;
Nitric Oxide
;
metabolism
;
Nitric Oxide Synthase
;
genetics
;
metabolism
;
Oryza
;
genetics
;
Plant Proteins
;
genetics
;
metabolism
;
Triticum
;
enzymology
;
genetics

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