Objective To observe the influence of environment stimulation on learning and memory ability and hippocampal pathology of neonatal rats with hypoxic-ischemic brain damage (HIBD).Methods The models of HIBD SD rats were established by the method of Rice, and were divided randomly into three groups: enriched environment stimulation group (EE), impoverished environment stimulation group (IE), and standard environment stimulation group (SE). The sham-operation rats were served as control group. Different environment stimulation was administrated to the rats since day 1 after HIBD.On the day 28,Morris water maze was used to evaluate the learning and memory ability. HE staining and nissl stain were employed to observe the pathological change and the number of neurons in hippocampus of rats.Results The learning and memory ability of EE group was significantly higher than that of SE group (P0.05), and the ability of SE group was higher than that of IE group ( P0.05). The number of SE group was lower than that of Sham group (P

Aminoacyl-tRNA syntheses (AARS) can catalyze the adenosine triphosphate (ATP)-dependent acylation of their cognate tRNA(s) with a specific amino acid. They can be seen as an index to reflect the energy metabolic rate of ischemic brain cells in ischemic penumbra. This study examined the relationship between arginyl-tRNA synthetase (ArgRS), one of the AARS, and cerebral ischemia in rats. The model of middle cerebral artery occlusion (MCAO) was established in rats. The expression levels of ArgRS protein and mRNA were detected in rat brain tissues at different time points following MCAO by Western blotting and RT-PCR, respectively. The results showed that the MCAO model was successfully established. Western blotting and RT-PCR analysis revealed that the ArgRS protein and mRNA were expressed in brain cells in both ischemic and normal penumbra tissues. The expression levels of ArgRS protein and mRNA peaked at 6 h after MCAO and decreased gradually. At 24 h, the expression levels of ArgRs protein and mRNA in ischemic penumbral tissues were lower than those in normal tissues. The expression levels of ArgRS mRNA and protein in ischemic penumbra varied with ischemic time, suggesting that the energy metabolism of brain cells in penumbra changed dynamically after ischemia to ensure the endogenous self-protection of the body. The brain oxygen supply should be improved as soon as possible, especially within 6-12 h after ischemia, so as to meet the demand for energy metabolism in ischemic penumbra and make sure the cell structure remains stable.

Morphology and molecular identification technology were used to identify 3 original plants of Fructus Elaeagni which was commonly used in Uygur medicine. Leaves, flowers and fruits from different areas were selected randomly for morphology research. ITS2 sequence as DNA barcode was used to identify 17 samples of Fructus Elaeagni. The genetic distances were computed by kimura 2-parameter (K2P) model, and the Neighbor-Joining (NJ) and Maximum Likelihood phylogenetic trees were constructed using MEGA5.0. The results showed that Elaeagnus angustifolia, E. oxycarpa and E. angustifolia var. orientalis cannot be distinguished by morphological characteristics of leaves, flowers and fruits. The sequence length of ITS2 ranged from 220 to 223 bp, the average GC content was 61.9%. The haplotype numbers of E. angustifolia, E. oxycarpa and E. angustifolia var. orientals were 4, 3, 3, respectively. The results from the NJ tree and ML tree showed that the 3 original species of Fructus Elaeagni cannot be distinguished obviously. Therefore, 3 species maybe have the same origin, and can be used as the original plant of Uygur medicineal material Fructus Elaeagni. However, further evidence of chemical components and pharmacological effect were needed.
DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Elaeagnaceae ; anatomy & histology ; classification ; genetics ; Fruit ; anatomy & histology ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Quality Control

Objective To explore the clinical application value of left atrial function with feature tracking cardiac magnetic resonance imaging (FT-CMR) by evaluating preliminarily left atrial strain and strain rate in patients with atrial fibrillation. Methods Thirty patients with paroxysmal atrial fibrillation, thirty patients with persistent atrial fibrillation and twenty-two healthy subjects were enrolled. All the subjects underwent cardiac magnetic resonance imaging with the real steady-state free precession(SSFP) sequence. FT-CMR parameters included left atrial strain and strain rate parameters, left atrial volume and function parameters were detected by using offline cardiovascular analysis software, respectively. Left atrial strain and strain rate parameters included left atrial total strain(Εs), passive strain(Εe), active strain(Εa), peak positive strain rate(SRs), peak early negative strain rate(SRe)and peak late negative strain rate(SRa). Volume and function parameters included maximum of left atrial volume(LAVmax), minimum of left atrial volume(LAVmin), total left atrial emptying fraction(LATEF), passive left atrial emptying fraction(LAPEF)and active left atrial emptying fraction(LAAEF). The differences in the general data among the paroxysmal atrial fibrillation group, the persistent atrial fibrillation group and the control group were compared by usingχ2 test or ANOVA analysis. The differences in all parameters between the atrial fibrillation group and the control group, the paroxysmal atrial fibrillation group and the persistent atrial fibrillation group were compared by using independent t test. Left atrial strain and strain rate parameters on an intra-observer and inter-observer were determined by intraclass correlation coefficient(ICC)analyses. Results Compared to control group, LAVmax and LAVmin in atrial fibrillation group were significantly increased(t=9.737,7.889,P<0.001);The LATEF and LAPEF had no significant difference, the LAAEF in two groups had statistically significant difference(t=-4.762,P<0.001).The absolute value of Es, Ee, Ea, SRs, SRe, SRa in atrial fibrillation group were significantly reduced than in control group(t=-7.732,-6.610,-6.493,-7.546, 6.864, 5.917,P<0.001). Compared with paroxysmal atrial fibrillation group, LAVmax and LAVmin in persistent atrial fibrillation group were increased obviously, LATEF and LAPEF were significantly decreased, and the differences were statistically significant(t=-4.575,-5.524, 4.002, 4.028,P<0.001).The LAAEF in two groups had no statistically significant difference. Compared with strain and strain rate in two groups, absolute value of Es, Ee, Ea, SRs, SRe, SRa in persistent atrial fibrillation group significantly decreased than in paroxysmal atrial fibrillation(t=4.310, 3.128, 4.465, 5.496,-3.290,-3.863,P<0.001). The intra-group and inter-group had well correlation coefficients between the observers in the left atrial strain and strain rate parameters of the subjects(ICC=0.85—0.94,0.81—0.90). Conclusions FT-CMR technique can be used to assess the left atrial strain and strain rate in patients with atrial fibrillation;Left atrial reservoir, conduit and booster-pump functions in patients with atrial fibrillation were impaired. Patients with persistent atrial fibrillation had worse left atrial function throughout the entire cardiac cycle compared with those with paroxysmal atrial fibrillation.

Objective Microenvironment plays important roles in the proliferation , viability, and apoptosis of tumor cells. This study was to investigate the effects of different functional groups on the biological characteristics of the osteosarcoma Saos -2 cell line in vitro. Methods Using self-assembled monolayers of alkanethiols on gold , we prepared different terminal chemical groups , including methyl (-CH3 ) , amino (-NH2 ) , hydroxyl (-OH) , and carboxyl (-COOH ) .We determined the similar density of different functional groups by contact angle measurement and x-ray photoelectron spectroscopy , and observed the effects of different functional groups on the adhesion , proliferation, viability, and apoptosis of the osteosarcoma Saos-2 cells by fluorescence microscopy , CCK-8 as-say, flow cytometry, and scan electron microscopy (SEM). Results The surface of -COOH and -NH2 promoted the adhesion and proliferation of the of the Saos-2 cells, with a good compatibility , while that of -CH3 was unfavorable for their adhesion and proliferation and even increased their apoptosis . The promoting effects of the functional groups on the adhesion and proliferation of the cells were listed in the following order: -COOH ≥ -NH2 >-OH -CH3 , while their toxicity and apoptosis-increasing effect ranked as -CH3 -OH >-NH2 >-COOH. Conclusion The-CH3 group inhibits the adhesion and proliferation and promotes the apoptosis of Saos-2 cells, which has provided some evidence for the surface design of biomaterials.

OBJECTIVE:To establish a method for simultaneous determination of liquiritin and glycyrrhizic acid in Wuwei shaji granules.METHODS:HPLC method was adopted.The determination was performed on Thermo scientific ODS-2 Hypersil column with mobile phase consisted of acetonitrile-0.5％ phosphoric acid (gradient elution) with flow rate of 1.0 mL/min.The detection wavelength was set at 237 nm,and column temperature was 25 ℃.The sample size was 20 μL.RESULTS:The linear ranges of liquiritin and glycyrrhizic acid were 48-480 μg(r=0.999 9) and 90-900 μg(r=0.999 9),respectively.The limits of quantitation were 0.55,2.10 μg/mL,and the limits of detection were 0.16,0.62 μg/mL.RSDs of precision,stability and reproducibility tests were all lower than 2.0％.The recoveries were 95.08％-97.58％ (RSD=0.93％,n=6)and 95.86％-99.89％ (RSD=1.67％,n=6),respectively.CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the content determination of liquiritin and glycyrrhizic acid in Wuwei shaji granules.

Aminoacyl-tRNA syntheses (AARS) can catalyze the adenosine triphosphate (ATP)-dependent acylation of their cognate tRNA(s) with a specific amino acid. They can be seen as an index to reflect the energy metabolic rate of ischemic brain cells in ischemic penumbra. This study examined the relationship between arginyl-tRNA synthetase (ArgRS), one of the AARS, and cerebral ischemia in rats. The model of middle cerebral artery occlusion (MCAO) was established in rats. The expression levels of ArgRS protein and mRNA were detected in rat brain tissues at different time points following MCAO by Western blotting and RT-PCR, respectively. The results showed that the MCAO model was successfully established. Western blotting and RT-PCR analysis revealed that the ArgRS protein and mRNA were expressed in brain cells in both ischemic and normal penumbra tissues. The expression levels of ArgRS protein and mRNA peaked at 6 h after MCAO and decreased gradually. At 24 h, the expression levels of ArgRs protein and mRNA in ischemic penumbral tissues were lower than those in normal tissues. The expression levels of ArgRS mRNA and protein in ischemic penumbra varied with ischemic time, suggesting that the energy metabolism of brain cells in penumbra changed dynamically after ischemia to ensure the endogenous self-protection of the body. The brain oxygen supply should be improved as soon as possible, especially within 6-12 h after ischemia, so as to meet the demand for energy metabolism in ischemic penumbra and make sure the cell structure remains stable.
Animals ; Arginine-tRNA Ligase ; biosynthesis ; Brain Ischemia ; genetics ; pathology ; Energy Metabolism ; Gene Expression Regulation ; Humans ; Oxygen Consumption ; RNA, Messenger ; biosynthesis ; Rats

OBJECTIVETo construct plasmid expressing pacemaker gene pIRES2-EGFP-HCN2 and study its effects in transfected atrial myocytes in vitro and in canine model of sick sinus syndrome (SSS).

METHODSmHCN2 gene was isolated from PTR plasmids and cloned into eukaryotic expression plasmid pIRES2-EGFP. Recombinant plasmids pIRES2-EGFP-HCN2 was transfected with by electroporation into neonatal atrial cardiomyocytes or injected to the sinoatrial (SA) region of canines with SSS induced by catheter and chemical ablation. pIRES2-EGFP-HCN2 expression was detected under fluorescence microscope and confirmed by reverse transcription-polymerase chain reaction (RT-PCR). Spontaneous beating rate in atrial cardiomyocytes was detected with light microscope.

RESULTSEGFP expression was seen in transfected atrial cardiomyocytes 24 to 48 hours after transfection and the spontaneous beating rate was significantly increased than that in non-transfected atrial cardiomyocytes [(180 +/- 11) bpm vs (140 +/- 14) bpm, P < 0.05]. Heart rate was significantly increased 24 hours post recombinant plasmids pIRES2-EGFP-HCN2 injection compared to saline injection in canines with SSS [(150 +/- 13) bpm vs (105 +/- 17) bpm, P < 0.05]. Green fluorescence was also detected in frozen SA tissue sections of canines injected with recombinant plasmids pIRES2-EGFP-HCN2 and the production amplified by RT-PCR was about 300 bp which is consistent with mHCN2 gene fragment.

CONCLUSIONThe recombinant eukaryotic expression plasmid pIRES2-EGFP-HCN2 can improve pacing function in atrial myocytes and in canine model of SSS.

Animals ; Disease Models, Animal ; Dogs ; Gene Expression ; Gene Transfer Techniques ; Genetic Therapy ; Genetic Vectors ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels ; In Vitro Techniques ; Ion Channels ; genetics ; Myocytes, Cardiac ; metabolism ; Plasmids ; Sick Sinus Syndrome ; therapy

Un-transfected rabbit corneal endothelial cells (RCECs) were cultivated, using chitosan blend membrane 4ha (chitosan-hyaluronic acid), 631ha (chitosan-hyaluronic acid) and 631s (chitosan-chondroitine sulfate) as scaffold carriers. Their biocompatibilities were studied in regard to cell adherence, morphological changes, growth status and monolayer forming abilities. The results indicated that RCECs cultivated on 4ha and 631ha carriers tended to be aggregated and even desquamated to some extent in local areas, and even more severely on 631ha carrier. And the RCECs cultivated on 631ha carrier could form almost a monolayer 48h later, and those on 4ha carrier could not. Contrarily, the RCECs cultivated on 631s carrier were evenly distributed and were in good status of growth with a good adherence and fibroblast-like morphology which could form almost a monolayer 48h later. And a complete monolayer was formed and was tightly attached to the 631s carrier 72h later. From the above results, it can be concluded that 631s carrier is most probably an ideal scaffold carrier for RCEC cultivation. 631s carrier may have the potential for use in the development of tissue-engineered rabbit corneal endothelium.
Animals ; Biocompatible Materials ; chemistry ; Cell Adhesion ; Cell Culture Techniques ; Cell Line ; Cells, Cultured ; Chitosan ; chemistry ; Endothelium, Corneal ; cytology ; Hyaluronic Acid ; chemistry ; Membranes, Artificial ; Rabbits ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

Objective To explore the effect of olfactory ensheathing cells (OECs) on the proliferation and differentiation of neural stem cells (NSCs). Methods The cells from the embryonic rat brain were primarily cultured and identified by immunofluorescence and immunocytochemistry. NSCs in the experimental group shared medium with OECs were cultured and induced to differentiate. Simultaneously, NSCs in the control group were cultured alone. The effect of OECs on the proliferation and differentiation of NSCs was observed by immunocytochemistry. Results Nerve growth factor receptor (P75NGFR) was observed in the primarily cultured OECs; nestin was expressed in the primarily cultured neurosphere and the cells differentiated from the neurosphere expressed neurofilament 200 (NF200) and glial fibrillary acidic protein (GFAP). Compared with that in the control group, the number of NSCs in the experimental group was significantly increased (P<0.05). On the 4~(th) and 7~(th) day of differentiation, the percentage of NF200-positive cell was higher in the experimental group than that in the control group (P<0.05), indicating that the appearance of OECs increased the differentiation of NSCs into NF200-positive cells. Conclusion OECs can promote the proliferation of NSCs and induce the differentiation of NSCs into neurons.