ObjectiveTo investigate the effect of soluble β-amyloid protein (Aβ) oligomers on the expression levels of insulin signaling transduction cascades-associated proteins including insulin receptor ( InsR),insulin receptor substrate-Ⅰ( IRS-Ⅰ) and protein kinase B (PKB) of rat hippocampal neurons,and the pathogenesis of Alzheimer's disease (AD) in depth.MethodsSoluble Aβ oligomers (5 μl) were injected into the lateral ventriculus of the AD group by a microinjector under the stereotaxic apparatus.Normal saline solution ( NS,5 μl) was injected into the NS group in the same way,and the control group received the puncture without injection. It was repeated after 1 week and the behavior of all rats was evaluatedbyY-mazetestafter2weeks.Thenhippocampuswasremovedandunderwent immunohistochemical staining to detect the expression of proteins associated.ResultsCompared with the other groups,learning and memory ability of the Aβ-treated rats were impaired.To be specific,the times of learning were increased and the times of memory were decreased. However,there was no significant difference between the NS group and the control group.Besides,the expression levels of InsR,IRS-Ⅰ,and PKB were decreased in AD group showing that a mean optical density of staining on these proteins ( InsR:0.12 ± 0.0l ; IRS-Ⅰ:0.14 ± 0.02; PKB:0.12 ± 0.03 ) was reduced in contrast with that in the NS group and the control group.Whereas there was no significant difference between the NS group (0.40 ± 0.02,0.39 ± 0.06,0.38 ± 0.03,mean difference:- 0.13,- 0.13,- 0.17,all P ＜ 0.05 ) and the control group (0.38 ± 0.07,0.35 ± 0.03,0.35 ± 0.06,mean difference:- 0.15,- 0.07,- 0.73,all P ＜ 0.05 ).ConclusionsSoluble Aβ1-42 induced learning and memory disability of the rats.The mechanism might be that Aβ can lead to disorders of the insulin signaling transduction pathway of hippocampal neurons and decrease the expression levels of the proteins in the pathway.

Objective To observe the change of endothelium-dependent vasodilatation function and to explore its mechanism in insulin-resistant (IR) rats induced by high fat feed. Methods (1) IR rat model was established by high fat feed for 4 weeks and IR was evaluated by glucose infusion rate (GIR) of euglycemic hyperinsulinemic clamp technique. (2) Acetylcholine (Ach)-dependent vasodilation response and nitric oxide synthase (NOS)-nitric oxide (NO)-cGMP function status were observed in isolated aorta of rats. Results (1) GIR was obviously lower in high fat feed group than that in routine feed group (P

Objective To study the development of neural stem cells from human fetal brains at different developmental stages. Methods A total of 100 cases of embryos at 16-32 gestational weeks by induction of labor with water bag were collected for the determination of the distribution, forms, existing modes, and the number of neural stem cells in the hippocampus by SABC immunohistochemical method and light microscopy. Results Neural stem cells were found in the hippocampus at different fetal ages and located in the polymorphic layer, pyramidal, granular and molecular layers of hippocampus, mainly in polymorphic layer, pyramidal layer, and granular layer. Neural stem cells in hippocampus were round, ellipse, triangle, and stellate, particularly round and ellipse. No obvious enation was found. Neural stem cells had plenty of cytoplasm. The nuclei were round and ellipse with rare chromatin and nucleoli from 2 to 6. Most of neural stem cells were distributed among other neurons, and symmetric cleavage was found in some of them, but some neural stem cells were distributed in cluster and nest. The number of neural stem cells in hippocampus were different between groups and gradually decreased with the increasing gestational age. Conclusion Neural stem cells exist widely in the hippocampus at different gestational ages. There are differences in distribution, forms, existing modes, and number of neural stem cells in hippocampus at different gestational ages. Hippocampus may be the new originating region of neural stem cells.

Objective: To prepare and characterize the polyclonal antibody against KIAA0649 and identify the localization and the functional motif of KIAA0649. Methods: Three polypeptides were synthesized based on the bioinformatics analysis of KIAA0649 protein. New Zealand rabbits were immunized with the mixture of the three KIAA0649 peptides coupled with keyhole limpet hemocyanin ( KLH). The titer of the antisera was detected with ELISA. The antisera were purified with immuno-affinity chromatography when the titer reached 1:10~5. Western blot was performed with the purified antisera on the cell lysates of U2OS cells transfected with either Flag-KIAA0649 or KIAA0649-targeting siRNA. Immunofluorescence was performed with the purified antisera and anti-Flag antibody on the cells transfected with FlagKIAA0649. A series of Flag-K1AA0649 deletion mutants was constructed by PCR cloning. The cellular compartmentation of full-length Flag-KIAA0649 and its deletion mutants were analyzed with immunofluorescence. Results: The results of Western blot and immunofluorescence demonstrated that the antisera from the KIAA0649 polypeptides-immunized rabbits specifically recognized endogenous and exogenous KIAA0649. The full-length Flag-K1AA0649 displayed specific nuclear foci. The Flag-KIAA0649 deletion mutant containing PENF motif showed the same nuclear foci as the full length of Flag-KJAA0649, suggesting that the PENF motif could be the minimum functional motif of KIAA0649. Conclusion: We have obtained anti-KIAA0649 polyclonal antibody which will be useful for further investigation. The PENF motifcould be the minimum functional domain of KIAA0649.

Objective To investigate the change of the content of polysaccharides in crude and processed Fructus Corni,thus to approach the process mechanism of Fructus Corni.Methods The content of polysaccharides in the crude and processed Fructus Corni was determined by phenol-sulfuric acid method.Results After processing with wine,the content of polysaccharides of Fructus Corni decreased from 10.12 %(content of total polysaccharidesbeing 51.41 %) to 5.91 %(content of total polysaccharidesbeing 53.10 %),decreased by 41.60 %as compared with that in the crude Fructus Corni.Conclusion After processing with wine,the content of polysaccharides decreases markedly.The results provide certain evidence for approaching the process mechanism of Fructus Corni.

OBJECTIVE:To study the chemical constituents of the ethyl acetate extraction portion of Polygonum bistorta,pro-vide scientific basis for the basic research of its effective substances. METHODS:Taking the ethyl acetate extraction portion of P. bistorta 95％ ethanol extract,positive phase and reversed phase column chromatography were used for repeated isolation and purifi-cation,and the structures of the purified compounds were identified on the basis of physicochemical properties or spectral data. RE-SULTS:11 monomer compounds were isolated and identified as β-sitosterol(1),dihydromyricetin(2),quercetin(3),kaempferol (4),epicatechin(5),chlorogenic acid(6),rutin(7),gallic acid(8),1,2,3,4-tetrahydro-8-hydroxy-4-isopropyl-1-methylnaph-thalene-6-carboxylic acid (9),protocatechuic acid (10) and ellagic acid (11). CONCLUSIONS:The literature is confirmed that compound 2 is isolated from P. bistorta for the first time and compound 9 is isolated from Polygonum for the first time. The related research results have provided experimental references for further clarifying the effective substances of P. bistorta.

Objective To investigate the role of Furin in breast cancer cell proliferation and provide a theoretical basis for in￣depth study of breast cancer. Methods Different concentrations of Furin inhibitor were added in MCF￣7 cell culture to test MCF￣7 cell proliferation by MTT essay.Hochest 33342 staining was used to detect the morphological change of apoptosic cells.Western blot analysis was applied to measure the level of cell apoptosis associated proteins,such as Caspase￣3,Caspase￣8 andCaspase￣9.The enzyme￣linked immunosorbent assay was used for detection the CAT and SOD levels in cell culture. ResultsMCF￣7 cell growth was inhibited by Furin inhibitor in a time and dose dependent manner.The results of Western blot and Hochest33342 staining indicated that MCF￣7 cells were apoptosis after Furin inhibitor treatment. The level of CAT was increasedsignificantly,associated with the level of SOD. Conclusion Furin inhibitor could induce MCF cell apoptosis, thereby inhibitcell proliferation by modulating MCF￣7 cell redox state.

Objective To evaluate the role of nuclear factor E2-related factor 2 (Nrf2)-antioxidant response element (ARE) signaling pathway in mitigation of anoxia/reoxygenation (A/R)-induced damage to rat cardiomyocytes by emulsified isoflurane postconditioning.Methods Primarily cultured cardiomyocytes obtained from Sprague-Dawley rats,aged 16-20 weeks,were seeded into the laminin pre-treated 6-well plates at a density of 104/cm2 and cultured for 20 h.The cells were randomly divided into 3 groups (n =12 each) using a random number table:control group (group C),group A/R,and emulsified isoflurane postconditioning group (group EI).The cells were cultured for 110 min without any treatment in group C.The cells were exposed to 45 min anoxia followed by 60 min reoxygenation in A/R and EI groups.After 45 min of hypoxia,the cells were cultured with emulsified isoflurane 1.68 mmol/L for 5 min before onset of reoxygenation in group EI.At the end of reoxygenation,transmission electron microscope was used to observe the ultrastructure of myocardial cells,and mitochondrial function was scored.The mRNA and protein expression of Nrf2,heme oxygenase-1 (HO-1),superoxide dismutase 1 (SOD1),and quinone oxidoreductase 1 (NQO1) was detected by real-time PCR and Western blot.Laser scanning confocal microscopy was used to detect Nrf2 nuclear translocation at the end of incubation,and Nrf2 activity was recorded in the nucleus of myocardial cells.Results Compared with group C,the mitochondrial function score was significantly increased,the mRNA and protein expression of Nrf2,HO-1,SOD1 and NQO1 was down-regulated,and Nrf2 activity in the nucleus was enhanced in A/R and EI groups.Compared with group A/R,the mitochondrial function score was significantly decreased,the mRNA and protein expression of Nrf2,HO-1,SOD1 and NQO1 was up-regulated,and Nrf2 activity in the nucleus was enhanced in group EI.Conclusion The mechanism by which emulsified isoflurane postconditioning mitigates A/R-induced damage to rat cardiomyocytes is related to induced nuclear translocation of Nrf2 and activated Nrf2-ARE signaling pathway.

Objective To investigate the characteristics of cognitive impairment in patients with type 2 diabetes mellitus (T2DM),and to analyze the correlation of T2DM with its risk factors and serum insulin like growth factor-1 (IGF-1) levels Methods A total of 78 hospitalized patients with T2DM at our hospital from November 2011 to March 2012 were divided into the cognitive impairment group (n=39) and the non-cognitive impairment group (n=39) according to Montreal Cognitive Assessment (MoCA) scores,and general clinical data were collected.Levels of blood lipids,glycosylated hemoglobin (HbAlc),fasting blood glucose (FBG),fasting blood insulin (FBI) and other biochemical indicators were detected,insulin resistance index (HOMA-IR) scores were calculated,and serum IGF-1 levels were determined by the enzyme-linked immunosorbent assay (ELISA).Results The education levle was (8.94±4.13) years for the cognitive impairment group and (12.65[2.50) years for the non-cognitive impairment group,and there was a statistically significant difference between the two groups (P=0.004).HbAlc levels were (9.69 ± 1.25) and (7.96 ± 1.31) for the cognitive impairment group and the non-cognitive impairment group,respectively,and were statistically difference between the two groups (P =0.001).Serum IGF-1 levels were (122.60±11.56) mmol/L and (139.32±9.57) mmol/L in the cognitive impairment group and the non-cognitive impairment group,respectively,and had a statistically significant difference between the two groups (P =0.037).Additionally,compared with the non-cognitive impairment group,scores on visuospatial ability,naming,language,abstraction,delayed recall and orientation were lower in the cognitive impairment group (P＜[0.05 or 0.01).Moreover,MoCA scores were negatively correlated with TC,LDL-C,TG,HbAlc,FBI levels and HOMA IR (r=0.498,-0.411,0.414,-0.452,-0.449,-0.539,respectively,P＜0.05 for all),and positively correlated with education lcvcl and IGF 1 level (r=0.579 and 0.491,respectively,P＜0.05 for both) Conclusions Cognitive impairment caused by T2DM is prominent in visuospatial ability,language,memory and executive functions,and is closely related to poor education,poor glycemic control,dyslipidemia and insulin resistance.Furthermore,decreased serum IGF-1 levels might be a risk factor for diabetic cognitive impairment.

The validity and reasonableness of emotional data are the key issues in the cognitive affective computing research. Effects of the emotion recognition are decided by the quality of selected data directly. Therefore, it is an important part of affective computing research to build affective computing database with good performance, so that it is the hot spot of research in this field. In this paper, the performance of two classical cognitive affective computing databases, the Massachusetts Institute of Technology (MIT) cognitive affective computing database and Germany Augsburg University emotion recognition database were compared, their data structure and data types were compared respectively, and emotional recognition effect based on the data were studied comparatively. The results indicated that the analysis based on the physical parameters could get the effective emotional recognition, and would be a feasible method of pressure emotional evaluation. Because of the lack of stress emotional evaluation data based on the physiological parameters domestically, there is not a public stress emotional database. We hereby built a dataset for the stress evaluation towards the high stress group in colleges, candidates of postgraduates of Ph. D and master as the subjects. We then acquired their physiological parameters, and performed the pressure analysis based on this database. The results indicated that this dataset had a certain reference value for the stress evaluation, and we hope this research can provide a reference and support for emotion evaluation and analysis.
Affect ; Computing Methodologies ; Databases, Factual ; Emotions ; Humans ; Recognition (Psychology) ; Stress, Psychological