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1.
Front Neurol ; 13: 942597, 2022.
Article in English | MEDLINE | ID: mdl-36062002

ABSTRACT

Objective: This systematic review and meta-analysis aimed to comprehensively evaluate the effectiveness and safety of acupuncture for post-stroke spasticity. Methods: Nine electronic databases were searched from their inception to 6 June 2022, to identify randomized-controlled trials (RCTs) that investigated the effectiveness and safety of acupuncture for post-stroke spasticity. Two reviewers independently screened the studies, extracted the data, assessed the risk of bias. The reporting quality of interventions in controlled trials of acupuncture was evaluated using Revised Standards for Reporting Interventions in Clinical Trials of Acupuncture (STRICTA). The RevMan 5.4 and R 4.2.0 software were used for statistical analysis. Results: A total of 88 eligible studies were included, involving 6,431 individuals. The pooled data demonstrated that acupuncture combined with conventional rehabilitation (CR) was superior to CR in reducing the Modified Ashworth Scale (MAS) score (standardized mean difference [SMD] = -0.73; 95% CI = -0.83 to -0.63; I 2 = 65%; low certainty of evidence). The favorable results were also observed in comparisons of acupuncture vs. CR (SMD = -0.22, 95% CI = -0.36 to -0.07; I 2 = 49%; moderate certainty of evidence). Subgroup analysis showed that acupuncture treatment with a frequency of once or twice a day was more effective than CR. In addition, the antispasmodic effect of acupuncture treatment increased with more sessions. Four studies explicitly reported slight acupuncture-related adverse events. Conclusion: Acupuncture could be recommended as adjuvant therapy for spasticity after stroke. However, due to the high risk of bias and heterogeneity of the included studies, the effectiveness of acupuncture for post-stroke spasticity remains to be confirmed.

2.
Food Chem ; 378: 132058, 2022 Jun 01.
Article in English | MEDLINE | ID: mdl-35032805

ABSTRACT

Dianhong tea (DHT) is popular for its pleasant caramel-like aroma. In this study, the aroma profile of high-grade DHT have been studied using gas chromatography-mass spectrometry (GC-MS) and gas chromatography-olfactometry (GC-O) combined with headspace solid phase microextraction (HS-SPME). A total of 52 aroma-active compounds were identified by GC-O coupled with aroma extract dilution analysis (AEDA) and odor specific magnitude estimation (Osme). Among them, quantification of 21 aroma-active compounds indicated that the content of linalool (5928 µg/kg) was the highest in high-grade DHT, followed by phenylethanol (3923 µg/kg) and phenylacetaldehyde (1801 µg/kg). Sensory-directed aroma recombination and omission tests further verified that phenylacetaldehyde, linalool, geraniol and 3-ethyl-2,5-dimethylpyrazine were important contributors to the overall sensory characteristics of high-grade DHT which dominated mainly by floral, sweet and caramel-like odors. This work will provide a theoretical reference for comprehensively understanding the aroma characteristic of DHT.


Subject(s)
Odorants , Volatile Organic Compounds , Gas Chromatography-Mass Spectrometry , Odorants/analysis , Olfactometry , Tea , Volatile Organic Compounds/analysis
3.
Funct Plant Biol ; 49(1): 102-114, 2021 12.
Article in English | MEDLINE | ID: mdl-34794538

ABSTRACT

Although the role of WRKY transcription factors (TFs) in colour formation has been reported in several species, their function in potato (Solanum tuberosum L.) anthocyanin biosynthesis remains unclear. In this study, the potato WRKY gene StWRKY13 was isolated and characterised. Expression analysis revealed a significantly higher StWRKY13 expression in chromatic tubers than in yellow ones. Transient activation assays showed that StWRKY13 could enhance the role of StAN2 in promoting anthocyanin biosynthesis in tobacco (Nicotiana tabacum L.). Over-expressing the StWRKY13 gene promoted anthocyanin biosynthesis in potato tubers. Further investigations indicated that StWRKY13 could interact with the StCHS, StF3H, StDFR, and StANS gene promoters and significantly enhance their activities. Our findings showed that StWRKY13 could promote anthocyanin biosynthesis by activating StCHS, StF3H, StDFR, and StANS transcription in potato tubers, thereby supporting the theoretical basis for anthocyanins formation in coloured potato tubers.


Subject(s)
Solanum tuberosum , Anthocyanins , Gene Expression Regulation, Plant , Plant Tubers/genetics , Solanum tuberosum/genetics , Nicotiana
4.
Sci Total Environ ; 752: 142264, 2021 Jan 15.
Article in English | MEDLINE | ID: mdl-33207511

ABSTRACT

Green synthesis of silver nanoparticles (Ag NPs) by using plants extracts has provided an eco-friendly alternation for industry and agriculture application. Here, we prepared Ag NPs by using the cucumber leaves and rice husk extracts, and further assessed the antimicrobial activity and phytotoxicity of green synthesized Ag NPs (g-Ag NPs) comparing with chemically synthesized Ag NPs (chem-Ag NPs). The chem-Ag NPs had strong antibacterial activity on the growth of Escherichia coli, while g-Ag NPs by rice husks (gr-Ag NPs) exhibited long-term antibacterial effects. In terms of phytotoxicity, the chem-Ag NPs induced over-generation of ROS and activated plant antioxidant defense systems, thus resulting in the upregulation of MDA and Zn contents and downregulation of antioxidant capacity, carotenoid, globulin and Mo contents. However, g-Ag NPs significantly promoted cucumber photosynthesis by increasing chlorophyll contents. Besides, the green synthesized Ag NPs by cucumber extracts (gc-Ag NPs) increased protein contents and gr-Ag NPs stimulated the upregulation of Mn and the downregulation of Al, which were all positive effects. Overall, compared with chem-Ag NPs, g-Ag NPs exhibited long-tern antimicrobial properties and attenuated toxicity to plants, which could be used as potential nanopesticide or nanoscale growth regulator in agriculture.


Subject(s)
Anti-Infective Agents , Metal Nanoparticles , Anti-Bacterial Agents/toxicity , Green Chemistry Technology , Metal Nanoparticles/toxicity , Microbial Sensitivity Tests , Plant Extracts/toxicity , Silver/toxicity
5.
PLoS Comput Biol ; 16(12): e1008489, 2020 12.
Article in English | MEDLINE | ID: mdl-33382685

ABSTRACT

The spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus poses serious threats to the global public health and leads to worldwide crisis. No effective drug or vaccine is readily available. The viral RNA-dependent RNA polymerase (RdRp) is a promising therapeutic target. A hybrid drug screening procedure was proposed and applied to identify potential drug candidates targeting RdRp from 1906 approved drugs. Among the four selected market available drug candidates, Pralatrexate and Azithromycin were confirmed to effectively inhibit SARS-CoV-2 replication in vitro with EC50 values of 0.008µM and 9.453 µM, respectively. For the first time, our study discovered that Pralatrexate is able to potently inhibit SARS-CoV-2 replication with a stronger inhibitory activity than Remdesivir within the same experimental conditions. The paper demonstrates the feasibility of fast and accurate anti-viral drug screening for inhibitors of SARS-CoV-2 and provides potential therapeutic agents against COVID-19.


Subject(s)
Aminopterin/analogs & derivatives , Antiviral Agents/pharmacology , Drug Evaluation, Preclinical/methods , Drug Repositioning , RNA-Dependent RNA Polymerase/antagonists & inhibitors , SARS-CoV-2/physiology , Aminopterin/chemistry , Aminopterin/pharmacology , Animals , Azithromycin/chemistry , Azithromycin/pharmacology , Chlorocebus aethiops , Computer Simulation , Deep Learning , Molecular Dynamics Simulation , RNA-Dependent RNA Polymerase/chemistry , Vero Cells , Virus Replication/drug effects , COVID-19 Drug Treatment
6.
Molecules ; 25(23)2020 Nov 30.
Article in English | MEDLINE | ID: mdl-33266001

ABSTRACT

In order to broaden the application of potato pulp pectic polysaccharide (PPP) in stabilizing acidified milk drinks (AMDs) and investigate the stabilizing effect and physical properties of AMDs prepared with PPP, a comparative study was made among PPP, commercial high methoxyl pectin (HMP) and low methoxyl pectin (LMP). The zeta potential, rheology, particle size and serum separation of AMDs were evaluated after preparing with PPP, HMP and LMP, respectively. Results indicated that PPP led to lower serum separation than LMP (14.65% for AMDs prepared with 0.5% PPP compared to 25.05% for AMDs prepared with 0.5% LMP), but still higher than HMP (9.09% for AMDs prepared with 0.5% HMP). However, narrower particle size distribution and lower viscosity of AMDs was achieved by PPP than by LMP and HMP. PPP can electrostatically adsorb on the surface of casein and its abundant neutral sugar side chains would provide steric hindrance to prevent casein flocculation in AMDs. Our results might provide some new ideas for the application of PPP in improving the stability of AMDs.


Subject(s)
Food Handling/methods , Milk Proteins/chemistry , Milk/chemistry , Pectins/chemistry , Polysaccharides/chemistry , Protein Stability , Solanum tuberosum/chemistry , Animals , Hydrogen-Ion Concentration , Rheology , Viscosity
7.
Molecules ; 25(5)2020 Mar 03.
Article in English | MEDLINE | ID: mdl-32138212

ABSTRACT

Dietary intake of potato starch could induce a dramatic increase in blood glucose and is positively associated with chronic metabolic diseases (type II diabetes, cardiovascular disease, etc.). Grape seed proanthocyanidins (GSP) are known to decrease starch digestion by inhibiting digestive enzymes or changing the physicochemical properties of starch. In the present study, GSP were complexed with potato starch to prepare polyphenol-starch complexes. The physiochemical properties and digestibility of complexes were investigated by in vitro digestion model, X-ray diffraction, differential scanning calorimetry, rapid visco analyzer, Fourier transform infrared spectroscopy as well as texture profile analysis. Results indicated that the peak viscosity, breakdown, trough, and setback of the complexes disappeared, replaced by a special continuous increase in paste viscosity. The complexes showed a lower final viscosity and higher thermal stability with the increasing binding amount of GSP. GSP decreased the hardness of the complexes' gel significantly. FT-IR indicated that GSP might interact with potato starch through noncovalent forces. Additionally, the complexes also showed a higher content of slowly digestible starch and resistant starch than that of the native starch. Thus, we inferred that the addition of GSP could modify the digestibility of potato starch and be an optional way to modify the starch with lower digestion.


Subject(s)
Grape Seed Extract/chemistry , Proanthocyanidins/chemistry , Solanum tuberosum/chemistry , Starch/chemistry , Calorimetry, Differential Scanning , Spectroscopy, Fourier Transform Infrared , X-Ray Diffraction
8.
Medicine (Baltimore) ; 98(39): e17124, 2019 Sep.
Article in English | MEDLINE | ID: mdl-31574812

ABSTRACT

INTRODUCTION: Spasticity is the most common complication after stroke, which is the main obstacle in the recovery of motor function. Spasticity seriously affects the quality of life and brings a heavy burden to families and society. Acupuncture is an effective method for stroke. However, whether acupuncture is effective for poststroke spasticity is still unknown. The purpose of this systematic review (SR) is to evaluate the effectiveness and safety of acupuncture for poststroke spasticity. METHODS AND ANALYSIS: We will search the following databases from inception to July 2019: China Biology Medicine (CBM), China National Knowledge Infrastructure (CNKI), Wan Fang Data, the Chinese Science and Technology Periodical Database (VIP), PubMed, Embase, The Cochrane Library, and Web of Science. All relevant randomized controlled trials (RCTs) utilizing acupuncture for poststroke spasticity will be included. The primary outcome is the modified Ashworth scale. Secondary outcomes include composite spasticity scale, clinic spasticity index, electromyographic activity, Hoffmann reflex activity, or other spasticity-related outcomes. Study selection, data extraction, and quality assessment will be performed independently by 2 reviewers. Assessment of risk of bias and data synthesis will be conducted using Review Manager V5.3 software. ETHICS AND DISSEMINATION: The ethical approval is not required since SR is based on published studies. The results of this SR will be published in a peer-reviewed scientific journal according to the Preferred Reporting Item for Systematic Review and Meta-analysis (PRISMA) guidelines. PROSPERO REGISTRATION NUMBER: CRD42019129779.


Subject(s)
Acupuncture Therapy/methods , Muscle Spasticity/therapy , Stroke/complications , Humans , Meta-Analysis as Topic , Muscle Spasticity/etiology , Research Design , Systematic Reviews as Topic , Treatment Outcome
9.
Plant Physiol Biochem ; 139: 411-418, 2019 Jun.
Article in English | MEDLINE | ID: mdl-30981157

ABSTRACT

Potato tuber dormancy is critical for the postharvest quality. The supply of carbohydrates is considered as one of the important factors controlling the rate of potato tuber sprouting. Starch is the major carbohydrate reserve in potato tuber, but very little is known about the specific starch degrading enzymes responsible for controlling tuber dormancy and sprouting. In this study, we demonstrate that an α-amylase gene StAmy23 is involved in starch breakdown and regulation of tuber dormancy. Silencing of StAmy23 delayed tuber sprouting by one to two weeks compared with the control. This phenotype is accompanied by reduced levels of reducing sugars and elevated levels of malto-oligosaccharides in tuber cortex and pith tissue below the bud eye of StAmy23-deficient potato tubers. Changes in soluble sugars is accompanied by a slight variation of phytoglycogen structure and starch granule size. Our results suggest that StAmy23 may stimulate sprouting by hydrolyzing soluble phytoglycogen to ensure supply of sugars during tuber dormancy.


Subject(s)
Germination/physiology , Plant Proteins/metabolism , Plant Tubers/metabolism , Plant Tubers/physiology , Solanum tuberosum/metabolism , Solanum tuberosum/physiology , alpha-Amylases/metabolism , Gene Expression Regulation, Plant/genetics , Gene Expression Regulation, Plant/physiology , Germination/genetics , Plant Proteins/genetics , Plant Tubers/genetics , Solanum tuberosum/genetics , Starch/metabolism , Sucrose/metabolism , Sugars/metabolism , alpha-Amylases/genetics
10.
Biomed Pharmacother ; 112: 108616, 2019 Apr.
Article in English | MEDLINE | ID: mdl-30780102

ABSTRACT

Depression pertains to the category of "Emotional Diseases" in traditional Chinese medicine (TCM). Its clinical symptoms are similar to the manifestations of "lily disease" from the TCM classics Synopsis of the Golden Chamber written by Zhang Zhongjing in the Han Dynasty. Also in this book, Lily Bulb and Rehmannia Decoction (LBRD) is the formula for the treatment of "lily disease". The classical herbal formula LBRD is composed of two herbs lily bulb and fresh rehmannia juice, with the function of nourishing yin and replenishing heart and lung. It has been clinically applied to treat "lily disease" for two thousand years. In this review, we focused on recent evidence linking LBRD and depression extracting data from animal and clinical studies, summarizing the primitive dosage and producing area of genuine medicinal materials of LBRD, clinical application, pharmacological mechanism and the effective substance basis for the treatment of depression. In conclusion, we discussed existing problems and future perspective. This systematic review will seek to enhance our understanding about pharmacology mechanism, herb-prescribing and recipe-constructing, and the development of novel formula for depression treatments.


Subject(s)
Biomedical Research/methods , Depression/drug therapy , Drugs, Chinese Herbal/therapeutic use , Lilium , Medicine, Chinese Traditional/methods , Rehmannia , Biomedical Research/trends , Depression/psychology , Drug Compounding , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/isolation & purification , Humans , Medicine, Chinese Traditional/trends , Treatment Outcome
11.
Plant Physiol Biochem ; 136: 215-221, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30690278

ABSTRACT

Potato cold-induced sweetening (CIS) is a major drawback restricting potato process industry. Starch degradation and sucrose decomposition are considered to be the key pathways in potato CIS. Our previous study showed that the RING finger gene SbRFP1 could slow down starch degradation and the accumulation of reducing sugars (RS) through inhibiting amylase and invertase activity in cold-stored tubers. However, the regulation mechanism of SbRFP1 is not clear. In this paper, we first proved that SbRFP1 could promote starch synthesis and modify the shape of starch granules. By further yeast two hybrid, GST-pull down and inhibition of enzyme activity assays, we confirmed that SbRFP1 could slow down the transformation of starch to RS in tubers mainly through the inhibition of ß-amylase StBAM1 activity. SbRFP1 was also proved to possess E3 ubiquitin ligase activity by ubiquitination assay. Thus, SbRFP1 may regulate the accumulation of RS in cold-stored tubers by ubiquitination and degradation of StBAM1. Therefore, our study reveals the regulatory mechanism of SbRFP1 in the process of CIS and provides more powerful evidence for the effect of starch degradation on potato CIS.


Subject(s)
Plant Proteins/physiology , Plant Tubers/growth & development , RING Finger Domains/physiology , Solanum tuberosum/growth & development , Amylases/metabolism , Cold Temperature , Gene Expression Regulation, Plant/genetics , Gene Expression Regulation, Plant/physiology , Genes, Plant/genetics , Genes, Plant/physiology , Plant Proteins/genetics , RING Finger Domains/genetics , Starch/metabolism , Two-Hybrid System Techniques , Ubiquitin-Protein Ligases/metabolism
12.
J Tradit Chin Med ; 39(5): 599-608, 2019 10.
Article in English | MEDLINE | ID: mdl-32186109

ABSTRACT

OBJECTIVE: To evaluate the effectiveness and safety of moxibustion therapy in the treatment of lumbar disc herniation (LDH). METHODS: Four Chinese databases and three English databases were searched from their inception to April 2018.Randomized controlled trials (RCTs) were included if moxibustion was used as the sole treatment or as a part of combination therapy with other treatments in patients with LDH. Two reviewers independently extracted the data and assessed the methodological quality using the Cochrane criteria for the risk of bias. The Meta-analysis was performed using Review Manager 5.3 software. RESULTS: In total, 16 RCTs including 1186 patients with LDH were analyzed. The Meta-analysis showed favorable effects of moxibustion in combination with massage therapy on the visual analog scale score compared with massage therapy alone [mean difference (MD) = -1.32, 95% confidence interval (CI) (-2.12, -0.51), P = 0.001]. The subgroup Meta-analysis failed to show favorable effects of electro-acupuncture plus moxibustion on the efficacy rate compared with electro-acupuncture alone [relative risk (RR) = 1.06, 95% CI (0.98, 1.14), P = 0.15]. However, acupuncture or massage therapy plus moxibustion improved the efficacy rates compared with acupuncture or massage therapy alone [RR = 1.33, 95% CI (1.18, 1.49), P < 0.000 01] [χ2 = 2.76, P = 0.25, I2 = 27%], [RR = 1.15, 95% CI (1.06, 1.25), P = 0.001] [χ2 = 0.00, P = 0.95, I2 = 0%]. With respect to the Japanese Orthopaedic Association (JOA) scores, acupuncture or massage therapy in addition to moxibustion produced results different from those of acupuncture or massage therapy alone [MD = 5.58, 95% CI (4.15, 7.00), P < 0.000 01] [χ2 = 0.58, P = 0.45, I2 = 0%], [MD = 3.61, 95% CI (3.01, 4.21), P < 0.000 01].There were no significant differences in the JOA score for subjective symptoms, objective symptoms, daily living ability, and other parameters. In six RCTs, no adverse reactions occurred during moxibustion. CONCLUSION: Whether moxibustion is an effective intervention for LDH is unclear because of the small sample size of qualified RCTs and the high risk of bias. More high-quality RCTs that overcome the methodological shortcomings of the existing evidence are needed.


Subject(s)
Intervertebral Disc Degeneration/therapy , Intervertebral Disc Displacement/therapy , Moxibustion/adverse effects , Moxibustion/methods , Safety , Humans , Treatment Outcome
13.
J Exp Bot ; 68(9): 2317-2331, 2017 04 01.
Article in English | MEDLINE | ID: mdl-28369567

ABSTRACT

Cold-induced sweetening (CIS) in potato is detrimental to the quality of processed products. Conversion of starch to reducing sugars (RS) by amylases is considered one of the main pathways in CIS but is not well studied. The amylase genes StAmy23, StBAM1, and StBAM9 were studied for their functions in potato CIS. StAmy23 is localized in the cytoplasm, whereas StBAM1 and StBAM9 are targeted to the plastid stroma and starch granules, respectively. Genetic transformation of these amylases in potatoes by RNA interference showed that ß-amylase activity could be decreased in cold-stored tubers by silencing of StBAM1 and collective silencing of StBAM1 and StBAM9. However, StBAM9 silencing did not decrease ß-amylase activity. Silencing StBAM1 and StBAM9 caused starch accumulation and lower RS, which was more evident in simultaneously silenced lines, suggesting functional redundancy. Soluble starch content increased in RNAi-StBAM1 lines but decreased in RNAi-StBAM9 lines, suggesting that StBAM1 may regulate CIS by hydrolysing soluble starch and StBAM9 by directly acting on starch granules. Moreover, StBAM9 interacted with StBAM1 on the starch granules. StAmy23 silencing resulted in higher phytoglycogen and lower RS accumulation in cold-stored tubers, implying that StAmy23 regulates CIS by degrading cytosolic phytoglycogen. Our findings suggest that StAmy23, StBAM1, and StBAM9 function in potato CIS with varying levels of impact.


Subject(s)
Amylases/genetics , Food Storage , Plant Proteins/genetics , Solanum tuberosum/physiology , Starch/metabolism , Amylases/metabolism , Gene Silencing , Plant Proteins/metabolism , Plant Tubers/genetics , Plant Tubers/physiology , RNA Interference , Solanum tuberosum/genetics , Sugars/analysis
14.
Plant Physiol ; 168(4): 1807-19, 2015 Aug.
Article in English | MEDLINE | ID: mdl-26134163

ABSTRACT

Slowing down cold-induced sweetening (CIS) of potato (Solanum tuberosum) tubers is of economic importance for the potato industry to ensure high-quality products. The conversion of sucrose to reducing sugars by the acid invertase StvacINV1 is thought to be critical for CIS. Identification of the specific StvacINV1 inhibitor StInvInh2B and the α- and ß-subunits of the interacting protein SUCROSE NONFERMENTING1-RELATED PROTEIN KINASE from the wild potato species Solanum berthaultii (SbSnRK1) has led to speculation that invertase activity may be regulated via a posttranslational mechanism that remains to be elucidated. Using bimolecular fluorescence complementation assays, this study confirmed the protein complex by pairwise interactions. In vitro kinase assays and protein phosphorylation analysis revealed that phosphorylation of SbSnRK1α is causal for StvacINV1 activity and that its active form blocks the inhibition of StInvInh2B by SbSnRK1ß, whereas its inactive form restores the function of SbSnRK1ß that prevents StInvInh2B from repressing StvacINV1. Overexpression of SbSnRK1α in CIS-sensitive potato confirmed that SbSnRK1α has significant effects on acid invertase-associated sucrose degradation. A higher level of SbSnRK1α expression was accompanied by elevated SbSnRK1α phosphorylation, reduced acid invertase activity, a higher sucrose-hexose ratio, and improved chip color. Our results lend new insights into a subtle regulatory mode of invertase activity and provide a novel approach for potato CIS improvement.


Subject(s)
Plant Proteins/metabolism , Protein Serine-Threonine Kinases/metabolism , Solanum tuberosum/metabolism , beta-Fructofuranosidase/metabolism , Amino Acid Sequence , Base Sequence , Blotting, Western , Gene Expression Regulation, Plant , Hexoses/metabolism , Molecular Sequence Data , Phosphorylation , Phylogeny , Plant Proteins/genetics , Plant Tubers/genetics , Plant Tubers/metabolism , Plants, Genetically Modified , Protein Binding , Protein Serine-Threonine Kinases/classification , Protein Serine-Threonine Kinases/genetics , Reverse Transcriptase Polymerase Chain Reaction , Solanum tuberosum/genetics , Sucrose/metabolism , beta-Fructofuranosidase/antagonists & inhibitors , beta-Fructofuranosidase/genetics
15.
Plant Sci ; 235: 14-24, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25900562

ABSTRACT

Promoter activity is crucial for precise gene expression. Previously, a synthetic tuber-specific and cold-inducible promoter, pCL, containing a C-repeat/dehydration-responsive element (CRT/DRE) cassette and a tuber-specific fragment, was constructed in order to regulate cold-induced sweetening (CIS) in potatoes. However, the utility of pCL is limited due to its low activity. To improve its inducibility in response to low temperatures, we modified the CRT/DRE and flanking sequences. In particular, promoter activity was significantly improved by site-specific mutation of flanking sequences next to the core element (CCGAC) of CRT/DRE. We also inserted a modified CRT/DRE cassette into pCL; although this enhanced activity, it was not more effective than mutation of the flanking sequences. Indeed, up to 20-fold enhanced pCL activity could be achieved by replacing the CRT/DRE cassette in pCL with tandem repeats of two mutated CRT/DRE cassettes. This improvement was due to an enhanced affinity between the CRT/DRE cassette(s) and the StCBF1 transcription factor. Together, these data suggest that altering the structure of CRT/DRE can enhance CBF-related transcription complex formation and thus improve the activity of this cold-inducible promoter.


Subject(s)
Carbohydrate Metabolism/genetics , Cold Temperature , Gene Expression Regulation, Plant , Plant Tubers/metabolism , Promoter Regions, Genetic , Solanum tuberosum/genetics , Transcriptional Activation , Arabidopsis/genetics , Genes, Plant , Genetic Engineering/methods , Mutation , Plant Proteins/genetics , Plants, Genetically Modified , Response Elements , Solanum tuberosum/metabolism , Trans-Activators/genetics , Transcription Factors/genetics , Transcription, Genetic
16.
Plant Biotechnol J ; 12(7): 984-93, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24985879

ABSTRACT

Potato cold-induced sweetening (CIS) is critical for the postharvest quality of potato tubers. Starch degradation is considered to be one of the key pathways in the CIS process. However, the functions of the genes that encode enzymes related to starch degradation in CIS and the activity regulation of these enzymes have received less attention. A potato amylase inhibitor gene known as SbAI was cloned from the wild potato species Solanum berthaultii. This genetic transformation confirmed that in contrast to the SbAI suppression in CIS-resistant potatoes, overexpressing SbAI in CIS-sensitive potatoes resulted in less amylase activity and a lower rate of starch degradation accompanied by a lower reducing sugar (RS) content in cold-stored tubers. This finding suggested that the SbAI gene may play crucial roles in potato CIS by modulating the amylase activity. Further investigations indicated that pairwise protein-protein interactions occurred between SbAI and α-amylase StAmy23, ß-amylases StBAM1 and StBAM9. SbAI could inhibit the activities of both α-amylase and ß-amylase in potato tubers primarily by repressing StAmy23 and StBAM1, respectively. These findings provide the first evidence that SbAI is a key regulator of the amylases that confer starch degradation and RS accumulation in cold-stored potato tubers.


Subject(s)
Amylases/metabolism , Plant Proteins/genetics , Solanum tuberosum/genetics , Starch/metabolism , Amino Acid Sequence , Amylases/antagonists & inhibitors , Amylases/genetics , Base Sequence , Carbohydrate Metabolism , Cloning, Molecular , Cold Temperature , Molecular Sequence Data , Phylogeny , Plant Proteins/chemistry , Plant Proteins/physiology , RNA, Messenger/metabolism , Solanum tuberosum/enzymology , Solanum tuberosum/metabolism
17.
Article in English | MEDLINE | ID: mdl-24319485

ABSTRACT

Qiangli Tianma Duzhong capsule (TMDZ), a Chinese herbal drug, is clinically used to improve functional outcome in patients with ischemic stroke in China. This study was conducted to establish whether postischemic long-term treatment with TMDZ could reduce the loss of injured hemisphere and confer the improvements of neurological outcome in chronic survival of rats with 2 h middle cerebral artery occlusion (MCAO)/reperfusion brain injury and its primary mechanisms. We found that TMDZ (44.5, 89, or 178 mg/kg), administered per os 6 h after the onset of ischemia and for 28 consecutive days, significantly improved the behavior deficits, beginning on day 7, and further improved later. TMDZ treatment also markedly reduced the tissue loss of the injured hemisphere and improved histopathology. In the meantime, TMDZ treatment could improve hemorrheology and inhibit platelet aggregation. These results provide the first evidence that post-ischemic long-term treatment with TMDZ confers the improvements of neurological outcome and the loss of injured hemisphere in an animal ischemic stroke model, and its mechanisms might be associated with the improvements of hemorrheology and the inhibition of platelet aggregation.

18.
Plant Physiol Biochem ; 73: 237-44, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24161651

ABSTRACT

The activity of vacuolar invertase (VI) is vital to potato cold-induced sweetening (CIS). A post-translational regulation of VI activity has been proposed which involves invertase inhibitor (VIH), but the mechanism for the interaction between VI and VIH has not been fully understood. To identify the potential partners of VI and VIH, two cDNA libraries were respectively constructed from CIS-resistant wild potato species Solanum berthaultii and CIS-sensitive potato cultivar AC035-01 for the yeast two-hybrid analysis. The StvacINV1 (one of the potato VIs) and StInvInh2B (one of the potato VIHs), previously identified to be associated with potato CIS, were used as baits to screen the two libraries. Through positive selection and sequencing, 27 potential target proteins of StvacINV1 and eight of StInvInh2B were clarified. The Kunitz-type protein inhibitors were captured by StvacINV1 in both libraries and the interaction between them was confirmed by bimolecular fluorescence complementation assay in tobacco cells, reinforcing a fundamental interaction between VI and VIH. Notably, a sucrose non-fermenting-1-related protein kinase 1 was captured by both the baits, suggesting that a protein complex could be necessary for fine turning of the invertase activity. The target proteins clarified in present research provide a route to elucidate the mechanism by which the VI activity can be subtly modulated.


Subject(s)
Enzyme Inhibitors/metabolism , Gene Expression Regulation, Plant , Plant Tubers/metabolism , Protein Processing, Post-Translational , Solanum tuberosum/genetics , Sucrose/metabolism , beta-Fructofuranosidase/genetics , Cryopreservation , DNA, Complementary , Diet , Food Storage , Genes, Plant , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Tubers/enzymology , Protein Kinases/genetics , Protein Kinases/metabolism , Solanum tuberosum/enzymology , Solanum tuberosum/metabolism , Species Specificity , Taste , Nicotiana/metabolism , Vacuoles/metabolism , beta-Fructofuranosidase/antagonists & inhibitors , beta-Fructofuranosidase/metabolism
19.
Plant Physiol Biochem ; 71: 77-86, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23886924

ABSTRACT

The cold-inducible promoter is ideal for regulating ectopic gene expression in plants to cope with the cold stress. The promoters of two cold-regulated genes, cor15a and cor15b, were cloned from Arabidopsis thaliana and their strengths were assayed in potato and tobacco. Although the cis-element composition and cold-inducible property were similar between the two promoters, the cor15b promoter showed significantly higher activity than the cor15a promoter in both potato and tobacco. In order to elucidate the factors determining this discrepancy, cor15a and cor15b promoters were separately truncated from 5'-end to construct short promoters with similar size containing a single C-repeat/dehydration-responsive element (CRT/DRE). Subsequently, two synthetic promoters were constructed by swapping the flanking sequences of CRT/DRE in the truncated promoters. The promoter strength comparison demonstrated that the flanking sequence could affect the promoter strength. These findings provide a potential regulatory mechanism to control the promoter strength without impact on other properties.


Subject(s)
Arabidopsis/genetics , Nicotiana/genetics , Promoter Regions, Genetic/genetics , Solanum tuberosum/genetics , Cold Temperature , Gene Expression Regulation, Plant/genetics , Gene Expression Regulation, Plant/physiology
20.
Plant Physiol Biochem ; 69: 9-16, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23688776

ABSTRACT

Potato vacuolar acid invertase (StvacINV1) (ß-fructofuranosidase; EC 3.2.1.26) has been confirmed to play an important role in cold-induced sweetening of potato tubers. However, the transcriptional regulation mechanisms of StvacINV1 are largely unknown. In this study, the 5'-flanking sequence of StvacINV1 was cloned and the cis-acting elements were predicted. Histochemical assay showed that the StvacINV1 promoter governed ß-glucuronidase (GUS) expression in potato leaves, stems, roots and tubers. Quantitative analysis of GUS expression suggested that the activity of StvacINV1 promoter was suppressed by sucrose, glucose, fructose, and cold, while enhanced by indole-3-acetic acid (IAA), and gibberellic acid (GA3). Further deletion analysis clarified that the promoter regions from -118 to -551, -551 to -1021, and -1021 to -1521 were required for responding to sucrose/glucose, GA3, and IAA, respectively. These findings provide essential information regarding transcriptional regulation mechanisms of StvacINV1.


Subject(s)
Gibberellins/pharmacology , Glucose/pharmacology , Indoleacetic Acids/pharmacology , Promoter Regions, Genetic/genetics , Solanum tuberosum/enzymology , Sucrose/pharmacology , Vacuoles/enzymology , beta-Fructofuranosidase/genetics , Gene Expression Regulation, Plant/genetics , Genotype , Plant Leaves/drug effects , Plant Leaves/enzymology , Plant Roots/drug effects , Plant Roots/enzymology , Plant Stems/drug effects , Plant Stems/enzymology , Promoter Regions, Genetic/drug effects , Solanum tuberosum/drug effects
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