Alzheimer's disease (AD) is a leading cause of dementia in the elderly. Mitogen-activated protein kinase phosphatase 1 (MKP-1) plays a neuroprotective role in AD. However, the molecular mechanisms underlying the effects of MKP-1 on AD have not been extensively studied. MicroRNAs (miRNAs) regulate gene expression at the post-transcriptional level, thereby repressing mRNA translation. Here, we reported that the microRNA-429-3p (miR-429-3p) was significantly increased in the brain of APP23/PS45 AD model mice and N2A^APP AD model cells. We further found that miR-429-3p could downregulate MKP-1 expression by directly binding to its 3'-untranslated region (3' UTR). Inhibition of miR-429-3p by its antagomir (A-miR-429) restored the expression of MKP-1 to a control level and consequently reduced the amyloidogenic processing of APP and Aβ accumulation. More importantly, intranasal administration of A-miR-429 successfully ameliorated the deficits of hippocampal CA1 long-term potentiation and spatial learning and memory in AD model mice by suppressing extracellular signal-regulated kinase (ERK1/2)-mediated GluA1 hyperphosphorylation at Ser831 site, thereby increasing the surface expression of GluA1-containing α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs). Together, these results demonstrate that inhibiting miR-429-3p to upregulate MKP-1 effectively improves cognitive and synaptic functions in AD model mice, suggesting that miR-429/MKP-1 pathway may be a novel therapeutic target for AD treatment.

Objective To study the role and mechanisms of equilibrative nucleotide transporter 1(ENT1)on Alzheimer's disease(AD)by constructing ENT1 overexpression and knockdown plasmids.Methods Molecular cloning was used to construct the ENT1 overexpression(pAAV-ENT1-mCherry)and knockdown(pAAV-ENT1shRNA-ZsGreen)plasmids.The overexpression plasmids and the knockdown plasmids were transfected into N2A cells(mouse Neuro A2 cells)and N2A-APP cells(N2A cells stably expressing human APP695).The expression of ENT1 and inflammatory factors at mRNA and protein levels were detected by real-time qPCR and Western blotting,respectively,and the change in cell viability were measured with CCK-8 assay.Results Sequencing and real-time qPCR indicated that ENT1 overexpression and knockdown plasmids were successfully constructed.CCK-8 assay showed that ENT1 overexpression significantly reduced the cell survival rate within 24 h(P＜0.05),while its knockdown increased the cell survival rate(P＜0.01).Real-time qPCR displayed that overexpression of ENT1 enhanced the expression levels of inflammatory factors,such as IL-1β,TNF-α,C1q-a and C1q-b in N2A cells(P＜0.05),while ENT1 knockdown reversed the above changes in inflammatory factors in N2A-APP cells(P＜0.05).Conclusion Knockdown of ENT1 attenuates pathological changes in AD by reducing the inflammatory response.ENT1 may be a potential target in the pathological mechanism of AD.

Objective To investigate the role of protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1) in the pathogenic process of Alzheimer's disease (AD).Methods The expression of POMGnT1 gene was examined in AD model cells (N2a/amyloid-precursor protein (APP) 695swe,n=3) and N2a/wt cells (n=3) using real-time PCR and Western blotting.This expression was also examined in AD model mice (APP/PS1,n=3) and wild-type mice (n=3) using immunofluorescence staining.Amyloid β-protein (Aβ) were examined in POMGnT1-gene-knockout mice (n=3) and wild-type mice (n=3) using immunochemistRy.Results The expression of POMGnT1 gene decreased in mRNA and protein levels in N2a/APP695swe cells compared to N2a/wt cells (mRNA:0.80±0.02 vs 1.00,t=10.52,P＜0.01;protein:0.50±0.02 vs 1.31±0.04,t=18.64,P＜0.01).Immunofluorescence results showed the reduced expression of POMGnT1 protein in neurons of APP/PS1 mice.Immunochemistry results showed more Aβ deposits in POMGnT1-gene-knockout mice (2 months old:0.358±0.014 vs 0.048±0.001,t=22.58,P＜0.01;1 year old:0.266±0.004 vs 0.229±0.003,t=7.771,P=0.002).Conclusion These findings suggest POMGnT1 gene may play an important role in the pathogenic process of AD.

Starting from the characteristics of nervous system,breaking the traditional subject-centered teaching model,combining the nervous system-related basic and clinical courses,Chongqing Medical University has established a nervous-module organ-system integration curriculum since 2010.This new teaching model is introduced and practiced in 5-year outstanding medical class of Chongqing Medical University.The new integrated curriculum guides students to learn knowledge from points to surface,helps them to combine knowledge vertically and horizontally,and simplifies the duplicate teaching content,so as to promote students to develop coherent and innovative thinking mode.Integrated curriculum pattern is an inevitable trend in medical education reform.

Objective To examine the changes of Aβ expression and its related metabolic enzymes in the brains of AD and T2DM mice, so as to explore the possible mechanism of type 2 diabetes mellitus combined with AD.Methods Five-month-old APP/PS1 double transgenic mice, ob/ob mice and the wild-type control mice were employed in this study.Immunohistochemical (IHC) staining, Elisa and Western blot were used to detect SP, Aβ and its related metabolic enzymes.Results A certain number of SPs were observed in the cerebral cortex and hip-pocampus of APP/PS1 mice;SPs were occasionally observed in the cortex of ob/ob mice, while no SP appeared in wild-type mice.Aβ40 and Aβ42 levels were significantly increased in APP/PS1 and ob/ob mice brains as compared with controls (P<0.05), thought both Aβ40 and Aβ42 levels in AD mice were significantly higher than those of ob/ob mice (P<0.05).APP expression level was highest in APP/PS1 mice among 3 groups, and its expressed higher in ob/ob mice than that of control mice (P<0.05).BACE1 expression was notably increased in APP/PS1 and ob/ob mice as compared with control(P<0.05), however, it expressed higher in APP/PS1 mice than ob/ob mice (P<0.05).The expression of Aβ degradation enzyme IDE was reduced in APP/PS1 and ob/ob mice(P<0.05), while lowest in ob/ob mice.Conclusions Overexpression of Aβ may be one of main reasons for T2DM combined AD.

Objective: To study the chemical constituents in the roots of Aconitum iochanicum Ulbr.Methods: The air-dried roots of A.iochanicum were powdered and extracted by methanol with a percolation method.After removing the solvent under reduced pressure, the crude extract was dissolved in1.5% HCl solution, and then extracted by ecetic ether.The acidic solution was basified to pH 9.0 by NaOH (5%) and extracted with ethyl acetate to obtain crude alkaloidal extract after the removal of ethyl acetate.The compounds were isolated and purified by column chromatography and identified based on spectral analysis (1H-NMR, 13C-NMR and MS).Results: Totally 18 compounds were isolated from A.iochanicum and characterized as 14-O-acetylsachaconitine (1), franchetine (2), crassicaudine (3), indaconoitine (4), 14-benzoyl chasmanine (5), 14-O-acetyltalatisamine (6), talatisamine (7), chasmannine (8), crassicauline A (9), bikhaconine (10), 13,15-dideoxyaconitine (11), crassicautine (12), kongboensine (13), liljestrandisine (14), ludaconitine (15), 8-deacetyl-yunaconitine (16), yunaconitine (17) and ouvrardiantine (18).Conclusion: It''s the first time to study the chemical constituents of A.iochanicum, and 18 diterpenoid alkaloids are isolated.

Objective:To study the secondary metabolites of endophytic fungus Alternaria solani from Aconitum Tsaii. Methods:The endogenous fungui fermentation product was extracted by methanol followed by recycling methanol to obtain the extract, and the ex-tract was extracted by ethyl acetate to obtain the crude extract. The compounds were isolated and purified by column chromatography and identified based on the spectral analysis ( MS, 1 H-NMR and 13 C-NMR) . Results:Totally 12 compounds were isolated from A. so-lani and characterized as (22E, 24R)-Ergosta-4, 6, 8 (14), 22-tetraen-3-one (1), 1,3-Diolein (2), 5-(3', 3'-Dimethylallyloxy)-3-methoxy-4-methylphthalide (3), pseudomonas aeruginosa H2-sesquiterpene lactone (4), ergosterol peroxide (5), 7-dehydroxyl-zinni-ol (6), ergosterol-7, 22-diene-3β, 5,6-triol (7), 9-octadecenoic acid -2',3'-dihydroxy propyl ester (8), 8-O-Methyltalatisamine (9), Chasmanine (10), yunaconitine (11) and crassicauline A (12). Conclusion:Totally 12 compounds are isolated from the endo-phytic fungus Alternaria solani, and four of them are isolated for the first time.

Objective:To study the secondary metabolites of endophytic fungus Alternaria solani from Aconitum Tsaii. Methods:The endogenous fungui fermentation product was extracted by methanol followed by recycling methanol to obtain the extract, and the ex-tract was extracted by ethyl acetate to obtain the crude extract. The compounds were isolated and purified by column chromatography and identified based on the spectral analysis ( MS, 1 H-NMR and 13 C-NMR) . Results:Totally 12 compounds were isolated from A. so-lani and characterized as (22E, 24R)-Ergosta-4, 6, 8 (14), 22-tetraen-3-one (1), 1,3-Diolein (2), 5-(3', 3'-Dimethylallyloxy)-3-methoxy-4-methylphthalide (3), pseudomonas aeruginosa H2-sesquiterpene lactone (4), ergosterol peroxide (5), 7-dehydroxyl-zinni-ol (6), ergosterol-7, 22-diene-3β, 5,6-triol (7), 9-octadecenoic acid -2',3'-dihydroxy propyl ester (8), 8-O-Methyltalatisamine (9), Chasmanine (10), yunaconitine (11) and crassicauline A (12). Conclusion:Totally 12 compounds are isolated from the endo-phytic fungus Alternaria solani, and four of them are isolated for the first time.

Objective:To study the spinal cord injury, spinal cord transection and persistent placeholder damage on the influence of secondary neural cell apoptosis in rats.Methods: Select 60 healthy male Wistar rats, numbered after using the random number table method is divided into A (18,spinal cord contusion),B (18,spinal cord transection),C (18,continuous placeholder),D (6,control),E (6,the control group only) groups of five,were observed at the 1,4,7 D after 5 group of rats nerve cell apoptosis index, spinal cord tissue Bcl-2,the expression of Bax,caspase 3 protein.Results:A,B,C three groups of rats after building 1 d are gray and white matter positive markers, and the gray matter and white matter of three groups of rats nerve cell apoptosis index differences statistically significant ( P<0.05);4 d,7 d after building gray matter and white matter of three groups of rats tend to place increased ap-optotic cells in the spinal cord index ( P<0.05);in building 1,4,7 d group C after rat spinal cord grey matter and white matter of apoptotic cell index was significantly higher than that of group A and group B, group B were significantly higher in group A and the differences were statistically significant (P<0.05).1,4,7 d after building A,B,C,D,E five group rats the Bcl-2,Bax,caspase-3 protein expression differences were statistically significant (P<0.05),1,4,7 d after building A,B,C the Bcl-2 of three groups of rats, Bax,caspase-3 protein expression was significantly higher than that of group D and group E ( P<0.05).Conclusion: Secondary rats after spinal cord injury of nerve cells apoptosis,apoptosis time,severity,and damage type and severity.

Objective To investigate whether catalpol affects senile plaque formation and spatial learning and memory ability in the amyloid-β protein precursor/presenilin 1 (APP/PSI) double transgenic mice.Methods Three month-old APP/PS1 double transgenic mice were randomly divided into catalpoltreated and saline-treated groups (n =10),with C57 mice of the same age and genetic background as normal control group (n =10).The catalpol (in a dose of 5 mg · kg-1 · d-1) and the same amount of saline were peritoneally injected into Alzheimer' s disease (AD) model mice for 3 weeks.Immunohistochemical staining was performed to examine senile plaques in the brain of AD model mice,and Morris water maze was used to assess the spatial learning and memory abilities of the mice.Results Compared with the saline-treated AD model mice (6.0 ±0.6),the number of senile plaques of catalpol treated AD mice significantly decreased (2.3± 0.7; t =3.500,P =0.025); Mice in each groups had similar latency and path length to reach platform in visible platform test; In hidden platform test,catalpol-treated mice had a significant lesser latency and path length compared with saline-treated mice,furthermore,catalpol-treated mice had much more platform-crossing times (6.4 ± 0.8) than saline-treated mice (2.9 ± 0.4 ; t =5.592,P =0.001).Conclusion Catalpol can significantly decrease the senile plaque formation and improve the spatial learning and memory abilities of APP/PS1 double transgenic mice.