In this study, we analyzed the composition and content of 25 free amino acids in 32 batches of different forms of Cervi Cornu Pantotrichum(CCP; one-branched, two-branched, and three-branched) from 15 producing areas. The clustering analysis and orthogonal partial least squares discriminant analysis(OPLS-DA) were performed based on the content of 25 free amino acids. Potential differential metabolites were identified based on VIP value. The results showed that there were 25 free amino acids in CCP, and the average content of essential, non-essential, and total amino acids was 6.13, 32.99, and 39.12 mg·g~(-1), respectively. The clustering analysis and OPLS-DA demonstrated that 25 free amino acids had different content among the three forms of CCP, of which two-branched CCP samples were separately gathered into a group. Five differential components, including glutamic acid, tryptophan, ornithine, γ-aminobutyric acid, and hydroxylysine, were screened out as potential quality markers for the identification of different forms of CCP. This study provides a theoretical basis for the quality evaluation, processing, and utilization of different forms of CCP.
Amino Acids/analysis* ; Animals ; Cornus ; Deer ; Gastropoda ; Glutamic Acid

PURPOSE: To investigate the 3-bond and spatial connectivity of human brain metabolites by scalar coupling and dipolar nuclear Overhauser effect/enhancement (NOE) interaction through 2D- correlation spectroscopy (COSY) and 2D- NOE spectroscopy (NOESY) techniques. MATERIALS AND METHODS: All 2D experiments were performed on Bruker Avance 500 (11.8 T) with the zshield gradient triple resonance cryoprobe at 298 K. Human brain metabolites were prepared with 10% D2O. Two-dimensional spectra with 2048 data points contains 320 free induction decay (FID) averaging. Repetition delay was 2 sec. The Top Spin 2.0 software was used for post-processing. Total 7 metabolites such as N-acetyl aspartate (NAA), creatine (Cr), choline (Cho), glutamine (Gln), glutamate (Glu), myo-inositol (Ins), and lactate (Lac) were included for major target metabolites. RESULTS: Symmetrical 2D-COSY and 2D-NOESY spectra were successfully acquired: COSY cross peaks were observed in the only 1.0-4.5 ppm, however, NOESY cross peaks were observed in the 1.0-4.5 ppm and 7.9 ppm. From the result of the 2-D COSY data, cross peaks between the methyl protons (CH3(3)) at 1.33 ppm and methine proton (CH(2)) at 4.11 ppm were observed in Lac. Cross peaks between the methylene protons (CH2(3,H alpha)) at 2.50ppm and methylene protons (CH2,(3,HB)) at 2.70 ppm were observed in NAA. Cross peaks between the methine proton (CH(5)) at 3.27 ppm and the methine proton (CH(4,6)) at 3.59 ppm, between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(4,6)) at 3.59 ppm, and between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(2)) at 4.05 ppm were observed in Ins. From the result of 2-D NOESY data, cross peaks between the NH proton at 8.00 ppm and methyl protons (CH3) were observed in NAA. Cross peaks between the methyl protons (CH3(3)) at 1.33 ppm and methine proton (CH(2)) at 4.11 ppm were observed in Lac. Cross peaks between the methyl protons (CH3) at 3.03 ppm and methylene protons (CH2) at 3.93 ppm were observed in Cr. Cross peaks between the methylene protons (CH2(3)) at 2.11 ppm and methylene protons (CH2(4)) at 2.35 ppm, and between the methylene protons(CH2 (3)) at 2.11 ppm and methine proton (CH(2)) at 3.76 ppm were observed in Glu. Cross peaks between the methylene protons (CH2 (3)) at 2.14 ppm and methine proton (CH(2)) at 3.79 ppm were observed in Gln. Cross peaks between the methine proton (CH(5)) at 3.27 ppm and the methine proton (CH(4,6)) at 3.59 ppm, and between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(2)) at 4.05 ppm were observed in Ins. CONCLUSION: The present study demonstrated that in vitro 2D-COSY and NOESY represented the 3-bond and spatial connectivity of human brain metabolites by scalar coupling and dipolar NOE interaction. This study could aid in better understanding the interactions between human brain metabolites in vivo 2DCOSY study.
Aspartic Acid ; Brain ; Choline ; Creatine ; Glutamic Acid ; Glutamine ; Humans ; Lactic Acid ; Protons ; Spectrum Analysis

Water soluble extract (WSE) is an important index for the quality evaluation of Astragali Radix (AR). In this study, the WSE of the wild AR from Shanxi province (SX) and the cultivated AR from Gansu Province (GS) were compared. The WSEs of two types of AR were determined according to the appendix of Chinese pharmacopoeia. Then the WSEs were subjected to NMR analysis, and the obtained data were analyzed using HCA, PCA, OPLS-DA, microarray analysis, and Spearman rank analysis. In addition, the Pearson correlation of differential metabolites were also calculated. The results showed that the WSE content of GS-AR (37.80%) was higher than that of SX-AR (32.13%). The main constituent of WSE was sucrose, and other 18 compounds, including amino acids, organic acids, were also detected. Multivariate analysis revealed that SX-AR contained more choline, succinic acid, citric acid, glutamate, taurine and aspartate, while GS samples contained more sucrose, arginine and fumaric acid. In addition, the Pearson correlations between different metabolites of the two types of AR also showed apparent differences. The results suggested that the WSE of two types of AR differs not only in the content, but also in the chemical compositions. Thus, the cultivation way is important to the quality of AR. This study supplied a new method for the comparison of extract of herbal drugs.
Arginine ; analysis ; Aspartic Acid ; analysis ; Choline ; analysis ; Citric Acid ; analysis ; Drugs, Chinese Herbal ; analysis ; chemistry ; Fumarates ; analysis ; Glutamic Acid ; analysis ; Magnetic Resonance Spectroscopy ; Multivariate Analysis ; Phylogeography ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Succinic Acid ; analysis ; Sucrose ; analysis ; Taurine ; analysis

Animals ; Glutamic Acid ; analysis ; toxicity ; Guinea Pigs ; Microscopy, Immunoelectron ; Vestibule, Labyrinth ; chemistry

OBJECTIVETo explore the effects of 1,2-dichloroethane (1,2-DCE) on the behavior and the brain neurotransmitter levels in mice.

METHODSThirty mice were randomly divided into four groups, which were control group and groups of low, middle and high exposure (225, 450 and 900 mg/m3) to 1,2-DCE for 10 days (3.5 h a day) by inhalation. After the last exposure, the open field test was performed immediately. After exposure all mice were killed and the brain tissues were taken up rapidly. The levels of aspartate (Asp), glutamate (Glu) and gamma-aminobutyric acid (GABA) in the brain were detected by high performance liquid chromatography (HPLC).

RESULTSLevels of Asp and Glu in all exposure groups increased with doses. As compared to the control group, levels of Glu in all exposure groups increased significantly (P < 0.05). Levels of GABA in the low exposure group were significantly lower than those in control group, but those in the high exposure group were significantly higher than those in control group. The results of the open field test showed that effect of low exposure to 1,2-DCE on the behavior was stimulant, but the high exposure to 1,2-DCE inhibited behavior of exploration, excitement and sport.

CONCLUSIONSSubacute exposure to 1,2-DCE could result in the change of amino acid neurotransmitter content and ratio in the brain, thereby change the behavior of mice appeared, which might be the mechanism of neurotoxicity caused by 1,2-DCE in part.

Animals ; Aspartic Acid ; analysis ; Behavior, Animal ; drug effects ; Brain ; metabolism ; Ethylene Dichlorides ; toxicity ; Female ; Glutamic Acid ; analysis ; Mice ; Mice, Inbred Strains ; Neurotransmitter Agents ; metabolism ; gamma-Aminobutyric Acid ; analysis

Yi-Gan San (YGS), a traditional Chinese medicine for dementia-related symptoms, was previously fractionated. One active fraction, YGS40 exhibited a neuroprotective effect against glutamate-induced cytotoxicity. In the present study, high-performance liquid chromatography, coupled with diode-array detection and quadrupole time-of-flight mass spectrometry, was applied for the identification of its chemical constituents and for quantification studies. The chemical profile of YGS40 consisted of sixty-four identified or tentatively characterized compounds. The levels of the major marker compounds increased significantly in the mixed decoction compared with those in the single plant decoction. The results suggest the high precision of the analyses of most of the constituents in YGS40 and establish the quantitative variations of the major marker compounds between the single and mixed decoction processes.
Chromatography, Liquid ; Cytotoxins ; Drugs, Chinese Herbal ; analysis ; Glutamic Acid ; toxicity ; Mass Spectrometry ; Neuroprotective Agents ; analysis ; therapeutic use

BACKGROUND: Sand blasted titanium (Ti) is commonly used in designing endosseous dental implants due to its biocompatibility and ability to form bonds with bone tissues. However, titanium implants do not induce strong interactions with teeth bones. To increase strong interactions between Ti disk implants and teeth bones, the L-glutamic acid grafted hydroxyapatite nanorods (nHA) were immobilized on albumin modified Ti disk implants (Ti-Alb). METHODS: For modification of Ti disk implants by nHA, the L-glutamic acid grafted nHA was synthesized and then immobilized on albumin modified Ti disk implants. Fourier transformed infrared spectroscopy, X-ray photoelectron spectroscopy, scanning electron microscope; energy dispersive spectroscopy and confocal laser scanning microscopy were used to confirm the modification of Ti disk implants. The bioactivity of nHA-modified Ti disk implants was evaluated by seeding MC3T3-E1 cells on Ti-nHA implants. RESULTS: Characterization techniques have confirmed the successful modification of Ti disk implants by L-glutamic acid grafted nHA. The nHA-modified Ti disk implants have shown enhanced adhesion, proliferation and cytotoxicity of MC3T3-E1 cells in comparison to pristine Ti implants. CONCLUSION: The modification of Ti implants by L-glutamic acid grafted nHA has produced highly osteogenic Ti disk plants in comparison to pristine Ti disk implants due to the formation of bioactive surfaces by hydroxyapatite nano rods on Ti disk implants. Ti-nHA disk implants showed enhanced adhesion, proliferation, and MC3T3-E1 cells viability in comparison to pristine Ti disk implants. Thus nHA might be to be useful to enhance the osseointegration of Ti implants with teeth bones.
Bone and Bones ; Dental Implants* ; Durapatite* ; Fourier Analysis ; Glutamic Acid ; Microscopy, Confocal ; Nanotubes ; Osseointegration ; Photoelectron Spectroscopy ; Spectrum Analysis ; Titanium* ; Tooth ; Transplants

OBJECTIVETo research the content changes of excitatory neurotransmitter and inhibitory neurotransmitter in corpus striatum of rats after single-used borneol and combining it with diazepam in hope of comprehending the activity of borneol on central nervous system and to observe whether borneol could increase the penetration of other drugs into the brain.

METHODThe content of four amino acids neurotransmitters in corpus striatum of rats were sampled by brain microdialysis technology at different time after administration and were determined by RP-HPLC which involved pre-column derivation with orthophthaladehyde (OPA), using phosphate gradient elution and fluorescence detection to detect the content of excitatory neurotransmitter aspartate (Asp), glutamate (Glu) and inhibitory neurotransmitter glycine (Gly), gamma-aminobutyric acid (GABA) in standards and samples and carry on statistical analysis.

RESULTThe content of both Gly and GABA in corpus striatum of rats with borneol increased significantly, compared with diazepam group (P < 0.05), while Asp and Glu showed no significant difference.

CONCLUSIONBorneol can improve permeability of diazepam through BBB.

Animals ; Aspartic Acid ; analysis ; Blood-Brain Barrier ; Bornanes ; administration & dosage ; pharmacology ; Corpus Striatum ; chemistry ; drug effects ; Diazepam ; administration & dosage ; pharmacology ; Glutamic Acid ; analysis ; Glycine ; analysis ; Male ; Neurotransmitter Agents ; analysis ; Rats ; Rats, Sprague-Dawley ; gamma-Aminobutyric Acid ; analysis

OBJECTIVETo investigate the variety of vitro recovery of amino acids for microdialysis probe after different dialysis time in vivo.

METHODSProbes were dialyzed in the amino acids standard solutions with microdialysis system,amino acid standard solutions and the microdialysate of probe were detected by the method of precolumn derivation with HPLC-RF.

RESULTAfter using different time of probe made by regenerated cellulose membrane, the vitro recoveries of Asp, Glu and GABA were not completely same (Asp: F=19.669, P=0.000; Glu: F=103.955, P=0.000; GABA: F=3.454, P=0.040); while the vitro recovery of Tau had no obvious difference(F=2.001, P=0.152). After using 6 h in vivo, recovery remain percentage (RRP) of Asp, Glu,Tau and GABA was 64.34 %, 67.36%, 103.11 % and 98.23 %, respectively, the recoveries of Asp, Glu decreased obviously (Asp: P < 0.01,Glu: P <0.05). After using 12 h in vivo, the RRP of Asp, Glu, Tau and GABA was 43.44 %, 24.42%, 77.45 % and 67.36 %, respectively, the recoveries of Asp, Glu and GABA decreased obviously (Asp: P < 0.001, Glu: P < 0.001, GABA: P < 0.05). After using 24 h in vivo, the RRP of Asp, Glu,Tau and GABA was 36.26 %, 12.24 %, 89.48 % and 71.35 %, respectively, the recoveries of Asp, Glu, GABA decreased obviously (Asp: P < 0.0001, Glu: P < 0.0001, GABA: P < 0.01).

CONCLUSIONDialysis in vivo could lead to the decline of recovery of probe, the decline is more obvious after longer dialysis. So when making brain dialysis experiments, the use time of probe should not be too long. To improve the validity of data, some calibration should be made on the recoveries of probe.

Amino Acids ; analysis ; Animals ; Aspartic Acid ; analysis ; Brain Chemistry ; Chromatography, High Pressure Liquid ; methods ; Dialysis Solutions ; analysis ; Glutamic Acid ; analysis ; Microdialysis ; methods ; Rats ; Rats, Sprague-Dawley ; Time Factors ; gamma-Aminobutyric Acid ; analysis

OBJECTIVETo determine whether there are in vivo differences of metabolites levels in bilateral cortical masticatory area(CMA) of patients with sleep bruxism, compared with healthy controls using proton magnetic resonance spectroscopy(1H-MRS). Accordingly to explore if cortical control of the central jaw motor system is abnormal in sleep bruxism patients.

METHODSFifteen sleep bruxism patients and fifteen age- and gender-matched healthy controls underwent 1H-MRS of bilateral CMA using J-difference edited point-resolved spectroscopy sequence(MEGA-PRESS) technique. Levels of metabolites were quantified from the ratio of the metabolite integral to the unsuppressed water signal. Differences of levels of γ-aminobutyric acid(GABA), glutmate(Glu) and N-acetyl aspartate(NAA) in bilateral CMA between sleep bruxism patients and healthy controls were tested using two-way ANOVA.

RESULTSEdited spectra were successfully obtained from the bilateral CMA in all of the participants. Levels of GABA+, glutmate and NAA in right and left CMA in sleep bruxism patients were (2.45±0.48)×10(-3), (2.35±0.62)×10(-3), (10.65±1.84)×10(-3), (10.49±2.37)×10(-3), (10.70±3.61)×10(-3), and (11.26±4.01)×10(-3) respectively. In contrast, levels of GABA+, glutmate and NAA in right and left CMA in healthy controls were (2.63±0.68)×10(-3), (2.65±0.97)×10(-3), (11.19± 1.34)×10(-3), (10.58±3.14)×10(-3), (11.82±1.80)×10(-3), and (11.95±3.23)×10(-3). There were no differences in levels of GABA+(P=0.196), Glu(P=0.590), and NAA(P=0.292) between sleep bruxism patients and healthy controls, nor in inbilateral CMA(GABA+: P=0.837; Glu: P=0.510; NAA: P=0.628).

CONCLUSIONSThe results indicate the absence of any alteration of the cortical control of the central jaw motor system in the levels of GABA, Glu and NAA in patients with sleep bruxism.

Analysis of Variance ; Aspartic Acid ; analogs & derivatives ; analysis ; metabolism ; Case-Control Studies ; Glutamic Acid ; analysis ; metabolism ; Humans ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy ; methods ; Masticatory Muscles ; metabolism ; physiopathology ; Motor Neurons ; metabolism ; Sleep Bruxism ; metabolism ; physiopathology ; gamma-Aminobutyric Acid ; analysis ; metabolism