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Ergothioneine is a multifunctional physiological cytoprotector, with broad application in foods, beverage, medicine, cosmetics and so on. Biosynthesis is an increasingly favored method in the production of ergothioneine. This paper summarizes the new progress in the identification of key pathways, the mining of key enzymes, and the development of natural edible mushroom species and high-yield engineering strains for ergothioneine biosynthesis in recent years. Through this review, we aim to reveal the molecular mechanism of ergothioneine biosynthesis and then employ the methods of fermentation engineering, metabolic engineering, and synthetic biology to greatly increase the yield of ergothioneine.
Subject(s)
Antioxidants , Ergothioneine/metabolism , Fermentation , Metabolic EngineeringABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of curcumin on oligomer formation and mitochondrial ATP-sensitive potassium channels (mitoKATP) induced by overexpression or mutation of α-synuclein.</p><p><b>METHODS</b>Recombinant plasmids α-synuclein-pEGFP-A53T and α-synuclein-pEGFP-WT were transfected into PC12 cells by lipofectamin method, and intervened by application of curcumin (20 μmol/L) and 5-hydroxydecanoate (5-HD). Oligomer formation in the cultured cells was identified by Western blotting and Dot blotting. Cytotoxicity and apoptosis of the PC12 cells were measured by lactate dehydrogenase (LDH) and JC-1 assays. mitoKATP were identified by Western blotting and whole cell patch clamp.</p><p><b>RESULTS</b>Curcumin has significantly reduced the oligomer formation induced by overexpression or mutation of α-synuclein in the cultured cells. LDH has decreased by 36.3% and 23.5%, and red/green fluorescence ratio of JC-1 was increased respectively by 48.46% and 50.33% after application of curcumin (P<0.05). Protein expression of Kir6.2 has decreased and mitoKATP channel current has significantly increased (P<0.05).</p><p><b>CONCLUSION</b>Curcumin can inhibit α-synuclein gene overexpression or mutation induced α-synuclein oligomers formation. It may block apoptosis induced by wild-type overexpression or mutation of α-synuclein. By stabilizing mitochondrial membrane potential. Opening of mitoKATP channel may have been the initiating protective mechanism of apoptosis induced by wild-type overexpression or mutation of α-synuclein. Curcumin may antagonize above cytotoxicity through further opening the mitoKATP channel.</p>
Subject(s)
Animals , Humans , Rats , Apoptosis , Cell Line , Curcumin , Pharmacology , KATP Channels , Chemistry , Genetics , Metabolism , Mitochondria , Genetics , Metabolism , Mutation , PC12 Cells , Parkinson Disease , Drug Therapy , Genetics , Metabolism , alpha-Synuclein , GeneticsABSTRACT
OBJECTIVE:To explore the effect of dexmedetomidine with different doses combined with ropivacaine on the bra-chial plexus block. METHODS:Totally 90 patients with upper extremity surgery of brachial plexus block were randomly divided in-to test group,control group 1 and control group 2. Test group was treated with 0.375%ropivacaine 40 ml+dexmedetomidine 100μg, brachial plexus injection;control group 1 was treated with 0.375% ropivacaine 40 ml+dexmedetomidine 50 μg,brachial plexus in-jection;and control group 2 was treated with 0.375% ropivacaine 40 ml,brachial plexus injection. Onset time of sensory and motor nerve block,block duration,postoperative pain time for the first time and pain score,analgesia duration,quality of analgesia and muscle relaxant,before anesthesia induction(T0),plasma cortisol concentrations in nerve block 10 min (T1),the start of surgery (T2)and incidence of adverse reactions were observed. RESULTS:The onset time of sensory and motor nerve block and postopera-tive pain score for the first time in test group werecontrol group 1>control group 2,plasma cortisol concentrations at T1 and T2 were higher than T0,and T2>T1>T0,test group0.05). CONCLUSIONS:Ropivacaine combined with 100 μg dexmedetomidine can effectively im-prove brachial plexus block and shorten the onset time of sensory and motor nerve block,with good safety.
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This study was aimed to preliminarily evaluate the acute toxicity and anti-inflammatory effects of coffee residue extract. The test maximum tolerated dose was applied in mice by gavage to observe the acute toxicity of coffee residue extract. Mice acute inflammation model was induced by xylene and glacial acetic acid. The gavage administration of coffee residue extract (1.00, 2.00, 3.00 g?kg-1, in terms of crude drug) was given 7 days con-tinuously. The ear swelling rate and celiac capillary permeability were measured. The results showed that the ex-tract of coffee residue maximum tolerated dose in mice is 8 . 60 g?kg -1 ( in terms of crude drug ) . The coffee residue extract of 3 . 00 g?kg-1 is able to inhibit ear swelling induced by xylene in mice ( P < 0 . 05 ) and the ex-cessive celiac capillary permeability ( P < 0 . 05 ) . It was concluded that the coffee residue extract have certain an-ti-inflammation activities.
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BACKGROUND: Magnetic resonance diffusion tensor imaging (MRDTI) may non-wounded detect damage of fiber in white matter and becomes an effectively way to evaluate upper motor neuron(UMN) impairments.OBJECTIVE: To investigate the clinical significance of MRDTI on amyotrophic lateral selerosis(ALS).DESIGN: Case contrast observation.SETTING: Department of Neurology, Renmin Hospital of Wuhan University.PARTICIPANTS: Twenty ALS patients were selected from Department of Neurology, Renmin Hospital of Wuhan University from April to December 2005. There were 11 males and 9 females, and their ages ranged from 33 to 73 years with the mean age of (51±10) years. All subjects met the diagnostic criteria of ALS set by World Neurology League.Other 15 healthy subjects were collected as control group. There were 8 males and 7 females, and their ages ranged from 31 to 73 years with mean age of (50±11) years. All subjects provided the confirm consent.METHODS: Based on level of upper and lower motor neuron impairments, ALS patients were divided into UMN impairment group (n =16) and lower motor neuron group (n =4). Functional scores of ALS, illness developing velocity and pyramidal sign scores were performed, respectively. All subjects were scanned with DTI at axial view. Regions of interest [subcortical white matter of precentral gyrul and postcentral gyrul (Pre-CG/Post-CG), centrum semiovale and frontal white matter (CS/FWM), peripheral lateral cerebral ventricle, posterior limb of internal capsule (PIC), cerebral peduncle (CP), genu corpus callosum and splenium corpus callosum (GCC/SCC) and dorsal thalamus (DT)] were selected to measure fractional anisotropy (FA) and apparent diffusion coefficient(ADC).MAIN OUTCOME MEASURES: Correlations among FA, ADC, functional score of ALS, illness developing velocity and pyramidal sign scores.RESULTS: Twenty patients and 15 subjects in the control group were involved in the final analysis. ① FA was reduced and ADC increased in the posterior limb of the internal capsule in patients with UMN signs compared to healthy volunteers (t =3.452, 2.670; P ＜ 0.01, 0.05). Nonparametric tests revealed that there was a trend toward reduced FA in the posterior limb of the internal capsule in B group compared to controls (U =11, P =0.057). ② In UMN impairment group, FA in the posterior limb of the internal capsule was positively correlated with the ALS rating scale (r =0.577, P ＜0.05) and negatively correlated with pyramidalsign scores (r = -0.789, P ＜ 0.01 ),CONCLUSION : The impairment of pyramidal tracts can be noninvasively evaluated by diffusion tensor MR in vivo, thus providing useful information in diagnosing and further understanding MND.
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Objective To investigate the function of human glia maturation factor gamma to induce the proliferation and differentiation of neural stem cells into astrocytes.Methods The neural stem cells were isolated from cerebral cortex of 11-to 12-day embryoes of SD rats.After three passages,the cells could react with nestin antigen,which demonstrated that the cells were neural stem cells.The neural stem cells were divided into 4 groups,control group,FBS group FBS,GMFG 5 ?g/L group and 5 ?g/L of GMFG,GMFG 10 ?g/L group with medium and 10 ?g/L of GMFG.Results In the control group,the cells had no significant differentiation in two days and slight branches were observed at the fourth day.In FBS group,the cells differentiation into astrocytes within three days,which grew on the surface of the culture flasks.In GMFG group,the cells differentiated into astrocytes in one day,and the differentiation of cells from the 10 ?g/L group was more obvious than that of cells from the 5 ?g/L group.The cells from the 10 ?g/L group reacted weakly with GFAP antigen.Conclusion GMFG can induce proliferation and differentiation of neural stem cells into astrocytes effectively in vitro.
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BACKGROUND: Nicotine, which is a known central nervous system stimulant, appears to be the neuroprotective factor of Parkinson disease(PD). It has been reported that PD patients' symptoms such as trembling,rigor, hypokinesia are ameliorated during smoking, but its mechanism still keeps unclear.OBJECTIVE: To observe the effects of nicotine on gene expression levels of dopamine D1 and D2 receptors (D1R,D2R)in striatum of rats and analyze the possible mechanism of behavioral changes of rats induced by nicotine.DESIGN:Randomized and controlled experiment.SETTING:Institute of Neurology, Xiangya Hospital, Central South University.MATERIALS :Twenty-four SD rats aged at 10 weeks were chosen,weighing 180-200 g. Nicotine (Sigma),revert AidTM M-Mulv reverse transcriptase (MBI Fermentas,USA), polymerase chain reaction (RCR,Beckman),densitometric scanning imaging system (Stratagene Eagle Eye Ⅱ ,USA).METHODS :This experiment was carried out in the Laboratory of Institute of Neurology, Xiangya Hospital,Central South University from July 2001 to July 2002. These rats were divided into two groups: control group (n=12)and nicotine group(n=12). The level of D1 and D2 receptors on striatum of rats was estimated at the timepoint of thirty-minute after chronic nicotine administration (4 mg/kg per day s.c.), and the behavioral activities were also recorded at the same timepoint for thirty minutes. The functional behavioral activities recorded included: rearing up repeatedly, moving about, provoking, climbing, grooming, yawning, rotating, smelling and vomiting. At the fourteenth day, all rats were killed after thirty minutes of nicotine injection,the brains were dissected out and the region of striatum was separated immediately. Total RNA was extracted from striatum by RNeasy Total RNA Kit. PCR amplification was performed at special condition. For semi-quantitative analysis, 10 μ L of PCR products for each was examined by electrophoresis on 12 g/L agarose gel containing 0.5 mg/L ethidium bromide,and absorbance (A value) was quantitated by using densitometric scanning imaging system, thuse D1R,D2R mRNA expression were determined. Differences between means were analyzed with two-tailed student's t test.MAIN OUTCOME MEASURFS: Changes of locomotor activities and the gene mRNA expression levels of D1 R and D2R in the regions of striatum in rats.RESULTS: Totally 24 SD rats were involved in the final results.① Locomotor activities of rats become more active after 3-day nicotine administration and reach the top during 7-14 days.②The A value of total RNA ratio of A260/A280 ＞1.8, and the total RNA had no degradation with 12 g/L agarose gels electrophoresis. ③As expected, PCR amplification product lengths of D1R, D2R,βA were 350 bp, 399 bp, 218 bp respectively. A significant increase of 23% of D1R mRNA expression in the region of striatum detected in the nicotine group compared with that of control group (98.63±1.13 and 65.29±1.45 seperately,P ＜ 0.01), no difference was detected on the level of D2R mRNA expression in the same regions above (76.73±1.45 and 78.21±1.69 respectively ,P ＞ 0.05 ).CONCLUSION: Nicotine may induce changes of locomotor activities of rats by up-regulating D1R mRNA expression in striatum.
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Objective To explore the effects of microstructure grinded on the labial face of porcelain fused to metal crown (PFM) in esthetic prosthesis for anterior teeth. Methods 193 patients with single maxillary incisor tooth defects were repaired with 2 crowns for each one. After the crowns being blent colour and molded, the crown A was not grinded particularly on the labial face and the crown B was done according to the records of the microstructure.The patients were allowed to choose the most satisfied one when 2 crowns were cemented temporarily. Results 36 patients chose the crown A and 157 chose B.The difference was significant ( P0.05). Conclusion The crown has great vitality and accordes with the esthetic standards of the patients by way of being grinded microstructure on the labial face.