Objective To establish a off-pump leukocyte depletion method in canines.Methods Twenty-one healthy adult mongrel dogs of both sexes, weighing 10-12 kg, were randomized into 2 groups: control group (group C, n = 9) and leukocyte depletion group (group LD, n = 12) . An extracorporeal leukocyte filtration end was constructed by aseptic puncture of the bilateral external jugular veins via a blood transfusion line and the leukocyte filter for extracorporeal leukocyte depletion. In group LD, blood was filtered with a MP-300 blood line and a SQ40S leucocyte depletion filter one end placed in the right external jugular vein (artery end) and the other end placed in the left external jugular vein (venous end). After heparin anticoagulation, a MP-300 blood line pump was used as the power. Blood was filtered at a rate of 75 ml/min and it was maintained for 60 min. The artery end was then closed, normal saline injected into the closed circuit, the remaining blood pumped into the body, and then the venous end closed. In group C, aseptic puncture of the bilateral external jugular veins was performed. Arterial blood samples were taken immediately before leukodepletion (baseline, T_0 ) , at 10, 20, 30, 40, 50 and 60 min of leukodepletion (T_(1-6) , depletion period), and at 10, 20, 30, 40, 50, 60, 120, 180 and 270 min after leukodepletion (T_(7-15) , recovery period) for determination of blood routine. MAP, HR, RBC.Plt and body temperature were recorded at T_0 , T_6 , T_(12) and T_(15) . Results There were no significant difference in MAP and RBC between the two groups ( P > 0.05 ) . HR, body temperature, and Plt were significantly lower in group LD than in group C (P < 0.05) .The leukocyte concentration was lower during depletion period in group LD than in group C ( P < 0.05) , while there was no significant difference in leukocyte concentration during recovery period between the two groups (P > 0.05 ). Conclusion The off-pump leukocyte depletion method is successfully established and has exact efficacy with less adverse effects in this study.

Memantine is a non-competitive N-methyl-D-aspartate receptor (NMDAR) antagonist clinically approved for moderate-to-severe Alzheimer's disease (AD) to improve cognitive functions. There is no report about the proteomic alterations induced by memantine in AD mouse model yet. In this study, we investigated the protein profiles in the hippocampus and the cerebral cortex of AD-related transgenic mouse model (3×Tg-AD) treated with memantine. Mice (8-month) were treated with memantine (5 mg/kg/bid) for 4 months followed by behavioral and molecular evaluation. Using step-down passive avoidance (SDA) test, novel object recognition (NOR) test and Morris water maze (MWM) test, it was observed that memantine significantly improved learning and memory retention in 3xTg-AD mice. By using quantitative proteomic analysis, 3301 and 3140 proteins in the hippocampus and the cerebral cortex respectively were identified to be associated with AD abnormalities. In the hippocampus, memantine significantly altered the expression levels of 233 proteins, among which PCNT, ATAXIN2, TNIK, and NOL3 were up-regulated, and FLNA, MARK 2 and BRAF were down-regulated. In the cerebral cortex, memantine significantly altered the expression levels of 342 proteins, among which PCNT, PMPCB, CRK, and MBP were up-regulated, and DNM2, BRAF, TAGLN 2 and FRY1 were down-regulated. Further analysis with bioinformatics showed that memantine modulated biological pathways associated with cytoskeleton and ErbB signaling in the hippocampus, and modulated biological pathways associated with axon guidance, ribosome, cytoskeleton, calcium and MAPK signaling in the cerebral cortex. Our data indicate that memantine induces higher levels of proteomic alterations in the cerebral cortex than in the hippocampus, suggesting memantine affects various brain regions in different manners. Our study provides a novel view on the complexity of protein responses induced by memantine in the brain of AD.
Alzheimer Disease ; Animals ; Axons ; Brain ; Calcium ; Cerebral Cortex ; Cognition ; Computational Biology ; Cytoskeleton ; Hippocampus ; Learning ; Memantine ; Memory ; Mice ; Mice, Transgenic ; N-Methylaspartate ; Proteome ; Ribosomes ; Water

Incarvillea younghusbandii Sprague is a traditional tonic herb. The roots are used as herbal medicine for nourishing and strengthening, as well as treating postpartum milk deficiency and weakness. In this study, the chloroplast genome of I. younghusbandii was sequenced and assembled by the high-throughput sequencing technology. The sequence characteristics, sequence repeats, codon usage bias, phylogenetic relationships and estimated divergence time of I. younghusbandii were analyzed. The 159 323 bp sequence contained a large single copy (80 197 bp), a small single copy (9 030 bp) and two inverted repeat sequences (35 048 bp). It contained 120 genes, including 77 protein coding genes, 8 ribosomal RNA genes and 35 transfer RNA genes. AAA was the most frequent codon in the chloroplast coding sequence of I. younghusbandii. A total of 42 simple sequence repeats were identified in the chloroplast genome. Phylogenetic analysis revealed I. younghusbandii was mostly like its taxonomically close relative Incarvillea compacta. The divergence between I. younghusbandii and I. compacta was dated to 4.66 million years ago. This study was significant for the scientific conservation and development of resources related to I. compacta. It also provides a basic genetic resource for the subsequent species identification of the genus Incarvillea, and the population genetic diversity study of Bignoniaceae.
Phylogeny ; Molecular Sequence Annotation ; Genome, Chloroplast ; Sequence Analysis, DNA ; Whole Genome Sequencing