The physiological and bio-marker function of D-acidic amino acids is now becoming the hot topic on metabolomics study and new drug discovery. A fully automated two-dimensional high performance liquid chromatography (2D-HPLC) system was established by using monolithic ODS column as the first dimension column, acetonitrile-trifluoro acetic acid-water (9: 0. 05: 92, V/ V) as the mobile phase; micro Chiralpak QD-1-AX column as the enantiomer separation column, 10 mmol/ L citric acid in methanol-acetonitrile (50: 50, V/ V) as the mobile phase for the second dimension, 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) as the fluorometrical derivative reagent. The separation efficiency ( Rs > 2. 5), determination sensitivity ( LOD =1 fmol) of acidic amino acids enantiomers were higher than those of existing methods, and an online confirmation of the enantiomers amounts was also achieved using this system. The recoveries were around 97-104% , RSD values for intra-day and inter-day precision were less than 5% for the acidic amino acids enantiomers in the biological samples. Furthermore, by analyzing the aging model senescence accelerated mouse prone 1 (SAMP1) mice which have low immunocompetence, the amounts of D-aspartic acid in thymus and spleen were determined as (206±18) and (264±21) nmol/ g, respectively. It is the first time that an obvious trend of the increasement of D-aspartic acid (p<0. 01) was observed in thymus and spleen of SAMP1 mice compare to senescence accelerated mouse resistant 1 (SAMR1) mice.

Objective To research the effects of neural stem cells (NSCs) transplantation on the neurological function improvement and neural survive and axonal regeneration in traumatic rat brain. Methods NSCs were cultured in vitro, labeled with hochest and transplanted into rat brain injured area by weight-dropping. Neurological severity scores(NSS) tested the functions at the 0,3,7,14 day post-injury. Immunofluorescence was used to detect the expressions of NeuN cells and GAP-43. Results There was significant difference in NSS between NSCs transplantion group and brain injury group(4.38 ±0.74 vs 5.50 ± 1.07, P＜0.01) at 7th day. There were a significant increase in neural number (51.46 ± 3. 303 vs 42.83 ± 5. 401, P ＜ 0.01 ) ), and GAP-positive axons ( 13.3 ± 1.7 vs 8.7 ± 1.1, P＜0.01 ) in NSCs transplantion group than in control group. Conclusion NSCs have an effects on the neurological improvement in brain injured rat. This may be associated to the increase in number of the neurons and local axons.

Objective To observe olfactory ensheathing cells transplantation on brain injury recovery nerve function and to explore its mechanism. Methods After purification of the olfactory ensheathing cells cultured for NGFRp75 immunocytochemical identification and preparation of cell suspension for transplantation, some cells were pre-labeled for bservation of survival after transplantation. 48 adult SD rats were randomly divided into three groups:sham operation without injury group (A group),cerebral cortex motor area injury group (B group) , the same brain injury and the olfactory ensheathing cell transplantation group (C group). At postoperative day,3 d, 7 d,14 d the neurological severity score (NSS) of rats were assessed; 14 d after injury of brain tissues were taken for NeuN immunohistochemistry host the number of neurons change. The data were statistically analyzed using SPSS17.0 software. Results (1) Cultured olfactory ensheathing cells showed NCFRp75 positive,the positive rate was 90%. (2) 14 d after transplantation of nuclear fluorescence labeling of olfactory ensheathing cells survived well in the host body. (3) 14 d after NSS score of B group( 2.00 ± 0.53) and C group ( 1.25 ± 0.46) were significantly better than the B group (P<0.05). (4) NeuN positive cells in B group (39.2 ±7. 1) and C group(45, 8 ± 6.0) were significantly better than B group (P<0.05). Conclusions Olfactory ensheathing cell transplantation can promote the recovery of neurological function in rats brain injury,which may be related with olfactory ensheathing cells to promote neuron survival in the host.

An analytical methodology based on an O-[2-(methacryloyloxy)-ethylcarbamoyl]-10,11-dihydroquinidine (MQD)-silica hybrid monolithic column was developed for the enantioseparation of 9-fluorenylmethoxycarbonyl (FMOC) derivatized amino acids by nano-liquid chromatography. The mo-bile phase was optimized including the apparent pH, content of ACN, and concentration of the buffer to obtain a satisfactory enantioresolution performance. 27 FMOC derivatized amino acids including 19 protein and 8 non-protein amino acids were tested, and 19 out of them were enantiomerically discriminated obtaining baseline separation for 11 of them. Analytical characteristics of the method were evaluated for norvaline and tryptophan in terms of linearity, precision, accuracy, limits of detection (LOD) and quantitation (LOQ) showing good performance to be applied to the enantiomeric determination of these amino acids in dietary supplements. LOD and LOQ values were 9.3 and 31μM for norvaline en-antiomers and 7.5 and 25μM for tryptophan enantiomers, respectively. The contents of D-norvaline and D-tryptophan were below their respective LODs in all the analyzed samples. Quantitation of L-tryptophan and L-norvaline showed good agreement with the labeled contents except for one sample which did not show presence of L-norvaline, contrary to the label indication.

ObjectiveTo explore the effects of Jinyaodai on neurological behavior and Akt expression in the cortex of rats following spinal cord contusion injury.MethodsRats were randomly divided into control group,spinal cord contusion group and Jinyaodai group.The weight-drop device was employed to prepare the spinal cord injury(SCI) model.Jinyaodai was administrated every day by using a stomach tube.Rats were performed the BBB assessment,and the detection of Akt expression and count of Neun positive neurons in cortex following SCI.ResultsCompared with control group,deficit of motor function in hindlimbs was seen at 3 dpo following cord contusion,and partial functional recovery could be seen from 7 dpo to 1 m.Treatment of Jinyaodai greatly increased the BBB scores ( 14.1 ± 1.4 ) more than SCI group ( 7.8 ± 1.3 ) at 1 month (P ＜ 0.05 ) ; Simultaneously,compared with SCI rats,treatment of Jinyaodai significantly increased the expression of Akt (0.53 ± 0.05,0.68 ± 0.07,P ＜0.05 ) and the number of neurons ( 11 ± 2, 15 ± 1 ; P ＜ 0.05 ) in the lesion-induced cortex of rats.Conclusion Jinyaodai may play an essential roles in functional recovery after spinal cord injury,in which the underlying mechanism may be involved in the expression of Akt in cortex.

ObjectiveTo explore the effects of bone morrow stromal cells (BMSCs) on the neurological behavior of rats with traumatic brain injury (TBI).MethodsTwenty-four SD rats were randomly and equally divided into control group, TBI group and BMSC group. The weight-drop device was adapted to establish the TBI model. The injury severity and its outcome were evaluated by a set of criteria termed neurological severity score (NSS). Brain tissues were harvested at day 14 to observe the survival and migration of the transplanted cells.Bax expression was detected by RT-PCR. Results NSS was (12 ±3 ) points in the TBI group, significantly higher than (7 ± 1 ) points in the BMSC group (P ＜0.05). The transplanted BMSCs could survive and migrate. Moreover, BAX, a crucail apopotosis gene, was down-regulated to 0.9 ±0.1 in the BMSC group, compared with 1.1 ±0.2 in the TBI group (P ＜0.05). ConclusionsBMSC transplantation is available to improve the neurological function, as may be associated with the Bax.

ObjectiveTo explore the effects of hyperbaric oxygen on neurological behavior and vascular endothelial growth factor (VEGF) in rats with traumatic brain injury (TBI). MethodsThirty rats were randomly divided into three groups, ie, control group, TBI group ( a 50 g weight-drop device was employed and fell from 30 cm height to induce the injury) and hyperbaric oxygen group ( HBO group,treated with hyperbaric oxygen once per day for seven days after TBI), 10 rats per group. Neurological severity score (NSS) was used to evaluate the movement and balance impairment in all groups. Expression of VEGF was detected by means of immunocytochemical staining.ResultsAfter TBI, the rats presented different degrees of convulsions, paralysis and balance dysfunction. The NSS score was (5.6 ±1.1 ) points in the TBI group and (0.3 ± O. 1 ) in the control group, with statistical difference ( P ＜0.05). While NSS score was (3.7 ± O. 7) points in the HBO group, showing a significant decrease compared with that in the TBI group (P ＜ 0. O1 ). Immunohistochemical staining showed 15 ± 3 positive neurons of VEGF in the TBI group, significantly less than 27 ± 2 in the control group ( P ＜ 0.05 ). There were 21 ±2 positive neurons of VEGF in the HBO group, significantly less than 21 ±2 in the TBI group (P ＜0.05). Conclusion Hyperbaric oxygen may attenuate experimental traumatic brain injury by stimulating production of VEGF.

ObjectiveTo explore the effect of swimming on motor behavior and expression of brain derived neurotrophic factor(BDNF) and trkB in adrenal body.MethodsSD adult rats were divided into normal cotrol group and swimming group ( n=10 in each group).All rats in swimming group were subjected 6 weeks swimming for 1h each day.Motor performances including swimming speed and distance were recorded and expression of BDNF and trkB in adrenal body was measured.ResultsThere was a significant increase in swimming speed ( ( 157 ± 60) m/min) and distance ( (283.36 ±49.50)m) in swimming group,compared with control group ( (283± 60) m/min,( 156.92 ± 29) m) (P ＜ 0.05 ).Simultaneously,expression of BDNF in adrenal body had been significantly unregulated (0.93 ± 0.09 vs 0.56 ± 0.19 ) (P ＜ 0.05 ),while expression of trkB kept to be not changed.ConclusionSwimming increases notor ability in rats,and the possible mechanism may be related the upregulation of BDNF in adrenal body.

In this study,a fluorescent(FL)aptasensor was developed for on-site detection of live Salmonella typhimurium(S.T.)and Vibrio parahaemolyticus(V.P.).Complementary DNA(cDNA)of aptamer(Apt)-functionalized multicolor polyhedral oligomeric silsesquioxane-perovskite quantum dots(cDNA-POSS-PQDs)were used as encoded probes and combined with dual-stirring-bar-assisted signal amplification for pathogen quantification.In this system,bar 1 was labeled with the S.T.and V.P.Apts,and then bar 2 was functionalized with cDNA-POSS-PQDs.When S.T.and V.P.were introduced,pathogen-Apt complexes would form and be released into the supernatant from bar 1.Under agitation,the two complexes reached bar 2 and subsequently reacted with cDNA-POSS-PQDs,which were immobilized on MXene.Then,the encoded probes would be detached from bar 2 to generate FL signals in the supernatant.Notably,the pathogens can resume their free state and initiate next cycle.They swim between the two bars,and the FL signals can be gradually enhanced to maximum after several cycles.The FL signals from released encoded probes can be used to detect the analytes.In particular,live pathogens can be distinguished from dead ones by using an assay.The detection limits and linear range for S.T.and V.P.were 30 and 10 CFU/mL and 102-106 CFU/mL,respectively.Therefore,this assay has broad application potential for simultaneous on-site detection of various live pathogenic bacteria in water.

Zwitterionic sulfobetaine-based monolithic stationary phases have attracted increasing attention for their use in hydrophilic interaction chromatography.In this study,a novel hydrophilic polymeric monolith was fabricated through photo-initiated copolymerization of 3-(3-vinyl-1-imidazolio)-l-propanesulfonate(SBVI)with pentaerythritol triacrylate using methanol and tetrahydrofuran as the porogenic system.Notably,the duration for the preparation of this novel monolith was as little as 5 min,which was significantly shorter than that required for previously reported sulfobetaine-based monoliths prepared via conventional thermally initiated copolymerization.Moreover,these monoliths showed good morphology,permeability,porosity(62.4％),mechanical strength(over 15 MPa),column efficiency(51,230 plates/m),and reproducibility(relative standard deviations for all analytes were lower than 4.6％).Mechanistic studies indicated that strong hydrophilic and negative electrostatic interactions might be responsible for the retention of polar analytes on the zwitterionic SBVI-based monolith.In particular,the resulting monolith exhibited good anti-protein adhesion ability and low nonspecific protein adsorption.These excellent features seem to favor its application in bioanalysis.Therefore,the novel zwitterionic sulfobetaine-based monolith was successfully employed for the highly selective separation of small bioactive compounds and the efficient enrichment of N-glycopeptides from complex samples.In this study,we prepared a novel zwitterionic sulfobetaine-based monolith with good performance and developed a simpler and faster method for preparation of zwitterionic monoliths.