1.Effect of ACC oxidase gene AhACOs on salt tolerance of peanut.
Jianbin HUANG ; Wenjie ZHOU ; Lei FANG ; Mingming SUN ; Xin LI ; Jingjing LI ; Xiaoting LI ; Yanyan TANG ; Defeng JIANG ; Hong ZHU ; Jiongming SUI ; Lixian QIAO
Chinese Journal of Biotechnology 2023;39(2):603-613
ACC oxidase (ACO) is one of the key enzymes that catalyze the synthesis of ethylene. Ethylene is involved in salt stress response in plants, and salt stress seriously affects the yield of peanut. In this study, AhACO genes were cloned and their functions were investigated with the aim to explore the biological function of AhACOs in salt stress response, and to provide genetic resources for the breeding of salt-tolerant varieties of peanut. AhACO1 and AhACO2 were amplified from the cDNA of salt-tolerant peanut mutant M29, respectively, and cloned into the plant expression vector pCAMBIA super1300. The recombinant plasmid was transformed into Huayu22 by pollen tube injection mediated by Agrobacterium tumefaciens. After harvest, the small slice cotyledon was separated from the kernel, and the positive seeds were screened by PCR. The expression of AhACO genes was analyzed by qRT-PCR, and the ethylene release was detected by capillary column gas chromatography. Transgenic seeds were sowed and then irrigated with NaCl solution, and the phenotypic changes of 21-day-seedings were recorded. The results showed that the growth of transgenic plants were better than that of the control group Huayu 22 upon salt stress, and the relative content of chlorophyll SPAD value and net photosynthetic rate (Pn) of transgenic peanuts were higher than those of the control group. In addition, the ethylene production of AhACO1 and AhACO2 transgenic plants were 2.79 and 1.87 times higher than that of control peanut, respectively. These results showed that AhACO1 and AhACO2 could significantly improve the salt stress tolerance of transgenic peanut.
Salt Tolerance/genetics*
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Arachis/genetics*
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Plant Breeding
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Ethylenes/metabolism*
;
Plants, Genetically Modified/genetics*
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Gene Expression Regulation, Plant
;
Plant Proteins/genetics*
2.Commentary: polymer binding modules accelerate enzymatic degradation of poly(ethylene terephthalate).
Yi LU ; Ruizhi HAN ; Ulrich SCHWANEBERG ; Yu JI
Chinese Journal of Biotechnology 2023;39(5):1883-1888
The large scale production and indiscriminate use of plastics led to serious environmental pollution. To reduce the negative effects of plastics waste on the environment, an approach of enzymatic degradation was put forward to catalyze plastics degradation. Protein engineering strategies have been applied to improve the plastics degrading enzyme properties such as activity and thermal stability. In addition, polymer binding modules were found to accelerate the enzymatic degradation of plastics. In this article, we introduced a recent work published in Chem Catalysis, which studied the role of binding modules in enzymatic hydrolysis of poly(ethylene terephthalate) (PET) at high-solids loadings. Graham et al. found that binding modules accelerated PET enzymatic degradation at low PET loading (< 10 wt%) and the enhanced degradation cannot be observed at high PET loading (10 wt%-20 wt%). This work is beneficial for the industrial application of polymer binding modules in plastics degradation.
Polyethylene Terephthalates/metabolism*
;
Polymers
;
Plastics
;
Ethylenes
3.Advances in poly(ethylene terephthalate) hydrolases.
Zhiyi ZHAO ; Guoqiang ZHANG ; Kun LIU ; Shengying LI
Chinese Journal of Biotechnology 2023;39(5):1998-2014
Plastics have brought invaluable convenience to human life since it was firstly synthesized in the last century. However, the stable polymer structure of plastics led to the continuous accumulation of plastic wastes, which poses serious threats to the ecological environment and human health. Poly(ethylene terephthalate) (PET) is the most widely produced polyester plastics. Recent researches on PET hydrolases have shown great potential of enzymatic degradation and recycling of plastics. Meanwhile, the biodegradation pathway of PET has become a reference model for the biodegradation of other plastics. This review summarizes the sources of PET hydrolases and their degradation capacity, degradation mechanism of PET by the most representative PET hydrolase-IsPETase, and recently reported highly efficient degrading enzymes through enzyme engineering. The advances of PET hydrolases may facilitate the research on the degradation mechanism of PET and further exploration and engineering of efficient PET degradation enzymes.
Humans
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Hydrolases/metabolism*
;
Polyethylene Terephthalates/metabolism*
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Plastics/metabolism*
;
Ethylenes
4.Gene cloning, induction, and prokaryotic expression of a Sm14-3-3 protein from Salvia miltiorrhiza.
Chen-Jing SHI ; Shi-Wei WANG ; Jia-Ming PENG ; Hai-Yu XU
China Journal of Chinese Materia Medica 2022;47(18):4886-4894
14-3-3 proteins are important proteins in plants, as they regulate plant growth and development and the response to biotic or abiotic stresses. In this study, a 14-3-3 gene(GenBank accession: OM683281) was screened from the cDNA library of the medicinal species Salvia miltiorrhiza by yeast two-hybrid and cloned. The open reading frame(ORF) was 780 bp, encoding 259 amino a cids. Bioinformatics analysis predicted that the protein was a non-transmembrane protein with the molecular formula of C_(1287)H_(2046)N_(346)O_(422)S_9, relative molecular weight of 29.4 kDa, and no signal peptide. Homologous sequence alignment and phylogenetic tree analysis proved that the protein belonged to 14-3-3 family and had close genetic relationship with the 14-3-3 proteins from Arabidopsis thaliana, Oryza sativa, and Nicotiana tabacum. The 14-3-3 gene was ligated to the prokaryotic expression vector pGEX-4 T-1 and then transformed into Escherichia coli BL21 for the expression of recombinant protein. Real-time fluorescent quantitative PCR showed that the expression of this gene was different among roots, stems, leaves, and flowers of S. miltiorrhiza. To be specific, the highest expression was found in leaves, followed by stems, and the lowest expression was detected in flowers. S. miltiorrhiza plants were treated with 15% PEG(simulation of drought), and hormones salicylic acid, methyl jasmonate, and ethephon, respectively, and the expression of 14-3-3 gene peaked at the early stage of induction. Therefore, the gene can quickly respond to abiotic stresses such as drought and plant hormone treatments such as salicylic acid, jasmonic acid, and ethylene. This study lays the foundation for revealing the molecular mechanism of 14-3-3 protein regulating tanshinone biosynthesis and responding to biotic and abiotic stresses.
14-3-3 Proteins/metabolism*
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Amino Acid Sequence
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Cloning, Molecular
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Ethylenes/metabolism*
;
Gene Expression Regulation, Plant
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Hormones/metabolism*
;
Phylogeny
;
Plant Growth Regulators/pharmacology*
;
Plant Proteins/metabolism*
;
Recombinant Proteins/genetics*
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Salicylic Acid/metabolism*
;
Salvia miltiorrhiza/metabolism*
5.Two Cases of Ethylene Glycol Poisoning.
Jae Jin KIM ; Sung Hyun YUN ; Hyun Min JUNG ; Ji Hye KIM ; Seung Baik HAN ; Jun Sig KIM ; Jin Hui PAIK
Journal of The Korean Society of Clinical Toxicology 2013;11(1):36-40
Ethylene glycol poisoning is treated mainly by alcohol dehydrogenase inhibition therapy and hemodialysis. Early recognition and initiation of treatment is important because toxic metabolites increase over time by hepatic metabolism; however, there is no confirmative diagnostic tool in our clinical setting. Therefore, diagnosis is dependent on history, high anion gap acidosis, high osmolal gap, etc.. Diagnosis and treatment are delayed in cases where history taking is not possible, such as a mental changed patient. Authors report on two cases of ethylene glycol poisoning by contrasting clinical outcomes, demonstrating the importance of early diagnosis and treatment for achievement of a good outcome.
Achievement
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Acid-Base Equilibrium
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Acidosis
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Alcohol Dehydrogenase
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Early Diagnosis
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Ethylene Glycol
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Ethylenes
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Osmolar Concentration
;
Renal Dialysis
6.The effect of a diode laser and traditional irrigants on the bond strength of self-adhesive cement.
Ali Riza TUNCDEMIR ; Cihan YILDIRIM ; Erhan OZCAN ; Serdar POLAT
The Journal of Advanced Prosthodontics 2013;5(4):457-463
PURPOSE: The purpose of this study was to compare the effect of a diode laser and traditional irrigants on the bond strength of self-adhesive cement. MATERIALS AND METHODS: Fifty-five incisors extracted due to periodontal problems were used. All teeth were instrumented using a set of rotary root canal instruments. The post spaces were enlarged for a No.14 (diameter, 1.4 mm) Snowlight (Abrasive technology, OH, USA) glass fiber reinforced composite post with matching drill. The teeth were randomly divided into 5 experimental groups of 11 teeth each. The post spaces were treated with the followings: Group 1: 5 mL 0.9% physiological saline; Group 2: 5 mL 5.25% sodium hypochlorite; Group 3: 5 mL 17% ethylene diamine tetra acetic acid (EDTA), Group 4: 37% orthophosphoric acid and Group 5: Photodynamic diode laser irradiation for 1 minute after application of light-active dye solution. Snowlight posts were luted with self-adhesive resin cement. Each root was sectioned perpendicular to its long axis to create 1 mm thick specimens. The push-out bond strength test method was used to measure bond strength. One tooth from each group was processed for scanning electron microscopic analysis. RESULTS: Bond strength values were as follow: Group 1 = 4.15 MPa; Group 2 = 3.00 MPa; Group 3 = 4.45 MPa; Group 4 = 6.96 MPa; and Group 5 = 8.93 MPa. These values were analysed using one-way ANOVA and Tukey honestly significant difference test (P<.05). Significantly higher bond strength values were obtained with the diode laser and orthophosphoric acid (P<.05). There were no differences found between the other groups (P>.05). CONCLUSION: Orthophosphoric acid and EDTA were more effective methods for removing the smear layer than the diode laser. However, the diode laser and orthophosphoric acid were more effective at the cement dentin interface than the EDTA, Therefore, modifying the smear layer may be more effective when a self-adhesive system is used.
Acetic Acid
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Axis, Cervical Vertebra
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Characidae
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Dental Pulp Cavity
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Dentin
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Edetic Acid
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Ethylenes
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Glass
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Incisor
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Lasers, Semiconductor*
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Methods
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Phosphoric Acids
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Resin Cements
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Smear Layer
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Sodium Hypochlorite
;
Tooth
7.Diosmin Reduces Calcium Oxalate Deposition and Tissue Degeneration in Nephrolithiasis in Rats: A Stereological Study.
Ali NOORAFSHAN ; Saied KARBALAY-DOUST ; Fatemeh KARIMI
Korean Journal of Urology 2013;54(4):252-257
PURPOSE: Kidney stones (nephrolithiasis) are a widespread disease. Thus, blocking stone formation and finding new therapeutic methods is an important area of study. Diosmin (a major component of the bile) is known to have antioxidant as well as renoprotective effects. The present investigation aimed to evaluate the effect of diosmin on renal tissue protection in rats with ethylene glycol-induced nephrolithiasis. MATERIALS AND METHODS: The rats were randomly divided into three groups. Group one (control) did not receive any treatments. In groups two and three, nephrolithiasis was induced by 2.5% (V/V) ethylene glycol + 2.5% (W/V) ammonium chloride (2 mL/d). The second and the third groups received distilled water or diosmin (80 mg/kg/d) by gavage for 21 days. RESULTS: Stereological estimation of the renal structures revealed that the average volume of calcium oxalate (CaOx) in the nephrolithiasis+diosmin rats was -63% less than in the rats with untreated nephrolithiasis (p<0.01). The volume of the glomeruli, proximal and distal convoluted tubules, Henle's loop, collecting ducts, and vessels was reduced -32% to 58% after the induction of nephrolithiasis (p<0.001). In the nephrolithiasis+diosmin rats, on average, -70% to 96% of the glomeruli, proximal convoluted tubules, Henle's loop collecting ducts, and vessels remained intact (p<0.01). Degeneration of the cortical tissue was 5-fold that of the medulla. In the nephrolithiasis+diosmin rats, degeneration in the renal cortical tissue and medulla was reduced -70% and 44%, respectively, compared with that in the untreated nephrolithiasis group (p<0.01). CONCLUSIONS: Diosmin reduces CaOx deposition and the degeneration of glomeruli and tubules in a rat model of nephrolithiasis.
Ammonium Chloride
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Animals
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Calcium
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Calcium Oxalate
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Diosmin
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Ethylene Glycol
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Ethylenes
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Kidney Calculi
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Nephrolithiasis
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Rats
;
Water
8.Chemical constituents of Rabdosia japonica var. glaucocalyx and their anti-complementary activity.
Shi YAO ; Nai-Yu XU ; Chun-Jun CHU ; Jian ZHANG ; Dao-Feng CHEN
China Journal of Chinese Materia Medica 2013;38(2):199-203
To study the chemical constituents of Rabdosia japonica var. glaucocalyx and their anti-complementary activity on the basis of preliminary studies. Target isolation guided by anti-complementary activity test, compounds in the chloroform and n-butanol fractions were isolated and purified by silica gel and Sephadex LH-20 column chromatographies, and preparative HPLC. The structures were identified by various spectroscopic data including ESI-MS, 1H-NMR and 13C-NMR data. The compounds were evaluated for anti-complementary activity in vitro. Eleven compounds were isolated from the chloroform and n-butanol soluble fractions and identified as stigmasterol (1), stigmas-9 (11) -en-3-ol (2), glaucocalyxin D (3), kamebakaurin (4), maslinic acid (5), corosolic acid (6), minheryins I (7), diosmetin (8), caffeic acid ethylene ester (9), caffeic acid (10) and vitexin (11). Isoquercetrin, rutin, quercetin, 3-methylquercetin, luteolin, 7-methylluteolin, and apigenin which were isolated from the preliminary studies together with compounds 9 and 10 showed inhibition of the complement system by the classical pathway. Compounds 2, 4, 6-9 and 11 were obtained from this plant for the first time. Caffeic acid (10) showed the strongest activity in vitro with a CH50 value of 0.041 g x L(-1).
Animals
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Antioxidants
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pharmacology
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Caffeic Acids
;
pharmacology
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Chromatography
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Chromatography, High Pressure Liquid
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Complement Hemolytic Activity Assay
;
methods
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Complement Inactivating Agents
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chemistry
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pharmacology
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Cricetinae
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Drugs, Chinese Herbal
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chemistry
;
isolation & purification
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Erythrocytes
;
drug effects
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Esters
;
Ethylenes
;
pharmacology
;
Female
;
Isodon
;
chemistry
;
Magnetic Resonance Spectroscopy
;
Male
;
Plant Components, Aerial
;
chemistry
;
Plant Growth Regulators
;
pharmacology
;
Sheep
;
Spectrometry, Mass, Electrospray Ionization
9.Direct biosynthesis of ethylene.
Chinese Journal of Biotechnology 2013;29(10):1431-1440
Ethylene is the most widely used petrochemical feedstock globally. The development of bio-ethylene is essential due to limited fossil fuels and rising oil prices. Bio-ethylene is produced primarily by the dehydration of ethanol, but can alternatively be directly produced from ethylene biosynthesis pathways in plants, algae, or microorganisms by using cheap and renewable substrates. This review addressed the biosynthesis of ethylene in plants and microorganisms, the characterization of key enzymes, genetic engineering strategies for ethylene biosynthesis in microorganisms, and evaluated its perspective and successful cases toward the industrial application. The direct production of bio-ethylene from a biological process in situ is promising to supplement and even replace the petrochemical ethylene production.
Ethylenes
;
biosynthesis
;
Industrial Microbiology
;
methods
;
Metabolic Engineering
;
methods
;
Plants
;
genetics
;
metabolism
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Saccharomyces cerevisiae
;
metabolism
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Synechocystis
;
genetics
;
metabolism
;
Trichoderma
;
metabolism
10.Vitrification of mouse embryos using the thin plastic strip method.
Eun Kyung RYU ; Yong Soo HUR ; Ji Young ANN ; Ja Young MAENG ; Miji PARK ; Jeong Hyun PARK ; Jung YOON ; San Hyun YOON ; Chang Young HUR ; Won Don LEE ; Jin Ho LIM
Clinical and Experimental Reproductive Medicine 2012;39(4):153-160
OBJECTIVE: The aim of this study was to compare vitrification optimization of mouse embryos using electron microscopy (EM) grid, cryotop, and thin plastic strip (TPS) containers by evaluating developmental competence and apoptosis rates. METHODS: Mouse embryos were obtained from superovulated mice. Mouse cleavage-stage, expanded, hatching-stage, and hatched-stage embryos were cryopreserved in EM grid, cryotop, and TPS containers by vitrification in 15% ethylene glycol, 15% dimethylsulfoxide, 10 microg/mL Ficoll, and 0.65 M sucrose, and 20% serum substitute supplement (SSS) with basal medium, respectively. For the three groups in which the embryos were thawed in the EM grid, cryotop, and TPS containers, the thawing solution consisted of 0.25 M sucrose, 0.125 M sucrose, and 20% SSS with basal medium, respectively. Rates of survival, re-expansion, reaching the hatched stage, and apoptosis after thawing were compared among the three groups. RESULTS: Developmental competence after thawing of vitrified expanded and hatching-stage blastocysts using cryotop and TPS methods were significantly higher than survival using the EM grid (p<0.05). Also, apoptosis positive nuclei rates after thawing of vitrified expanded blastocysts using cryotop and TPS were significantly lower than when using the EM grid (p<0.05). CONCLUSION: The TPS vitrification method has the advantages of achieving a high developmental ability and effective preservation.
Animals
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Apoptosis
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Blastocyst
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Dimethyl Sulfoxide
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Embryonic Structures
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Ethylene Glycol
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Ethylenes
;
Ficoll
;
Mental Competency
;
Mice
;
Microscopy, Electron
;
Plastics
;
Sucrose
;
Vitrification

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