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AIM:To investigate the protective effects of astrocyte protein phosphatase 2A (PP2A) up-regula-tion on APP/PS1 double transgenic mice.METHODS:An eGFP-wtPP2A lentivirus with glial fiber acidic protein promoter was constructed to specifically increase PP2A expression in the astrocytes.The mice were divided into wild -type mice +vector virus group (Con), APP/PS1 transgenic mice +vector virus group (APP/PS1) and APP/PS1 transgenic mice +eGFP-wtPP2A lentivirus group (PP2A) by lateral ventricular injection of the lentivirus.Four weeks after injection of the vi-rus, the immunofluorescence of brain slices were used to detect the level of β-amyloid protein ( Aβ) .Golgi staining was used to detect the changes of dendritic spine density and morphology.Electron microscopy was applied to detect the thickness of postsynaptic density (PSD).The Morris water maze test was applied to examine the learning and memory abilities of the mice.RESULTS: Up-regulation of PP2A in the astrocytes attenuated Aβlevel increasing in APP/PS1 group.Up-regulation of PP2A in the astrocytes significantly attenuated both decreases in the dendritic spine density and the percentage of mushroom-like dendritic spines in the hippocampal CA3 region of APP/PS1 mice.Up-regulation of PP2A in the astrocytes significantly attenuated the reduced thickness of PSD in APP/PS1 group.Up-regulation of PP2A in the astro-cytes attenuated the escape latency extending in APP/PS1 group .CONCLUSION: Up-regulation of PP2A in the astro-cytes reduces AD-like pathological changes, and attenuates synaptic impairment, synaptic plasticity deficits and cognitive impairment in the APP/PS1 double transgenic mice.
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Objective To analyze the epidemiological characteristics of Klebsiella pneumoniae car-bapenemase(KPC)-producing Escherichia coli(E. coli)strains isolated in Hangzhou,China. Methods A total of 25 KPC-producing Escherichia coli strains were collected from four hospitals in Hangzhou from July 2012 to January 2014. Antibiotic susceptibility of the isolates to 22 common antimicrobial agents was deter-mined by using Kirby-Bauer(K-B)disk diffusion method. PCR analysis and gene sequencing were used for bla KPC gene screening. The modified Hodge test was performed to detect the production of carbapenemase. Pulsed-field gel electrophoresis(PFGE)and multi-locus sequence typing(MLST)were used for homology analysis. Results All of the 25 clinical isolates were confirmed to be KPC-producing E. coli strains,harbo-ring the blaKPC-2 gene. These KPC-producing isolates showed high drug resistance rates and were resistant to almost all β-lactam antibiotics. PFGE typing classified the 25 isolates into three main homologous clone groups,including clone group A(4 isolates),clone group B(5 isolates)and clone group C(2 isolates), and some single clones(14 isolates). MLST typing classified the isolates into eight ST types,including ST131(14 isolates),ST167(3 isolates),ST2003(3 isolates),ST410(1 isolate),ST457(1 isolate), ST1463(1 isolate),STnew1(1 isolate)and STnew2(1 isolate). The typing results of PFGE and MLST were consistent with each other. Conclusion The prevalent KPC-producing E. coli strains in Hangzhou, China were ST131 type,which were resistant to multiple antibiotics and had been detected in several hospi-tals. The epidemic of KPC-producing E. coli strain often occurred at some special wards,such as Intensive Care Unit(ICU)and emergency ICU.
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Objective To investigate the expressions of KLF2 mRNA and KLF4 mRNA in the acute lung injury (ALl) rats induced by lipopolysaccharide (LPS),and to analyze the correlation between KLF2,KLF4 and ALI.Methods A total of 100 SD rats were randomly divided into 2 groups:normal control group and LPS treated group,then the latter group was randomly further divided into 3 subgroups as per the serum and lung tissue samples taken separately at 2,4 and 24h after modeling.The ALI model was made by injecting 5mg/kg LPS into tail vein.The pathological changes of lung tissue were observed in each group,and the expressions of KLF2,KLF4 mRNA in serum and lung tissue were detected by RT-PCR.The data of laboratory findings were analyzed with SPSS 17.0 software for statistical analysis.Results The histopathological changes showed the most obvious damage of lung tissue occurred at 4 hours after modeling.The expressions of KLF2 mRNA and KLF4 mRNA in the lung tissue and serum of control group were significantly higher compared to LPS treated subgroups (P ＜0.01).The expression of KLF2 mRNA in LPS treated subgroup at 2 hours was lower than that in LPS subgroups at 4 hours and 24 hours (P ＜ 0.01),while the expression of KLF4 mRNA in LPS treated subgroup at 4 hours was lower than that in LPS treated subgroups at 2 hours and 24 hours (P ＜ 0.01).Conclusions The expression of KLF2 mRNA was occurred earlier than the pathological changes in acute lung injury,while the expression of KLF4 was emerged synchronously,and both KLF2 and KLF4 could be used as candidates of predictive and diagnostics molecular markers of ALI.
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ObjectiveTo observe the apoptotic effect of cardamonin on K562 cells and its relationship with the expressions of PTEN, p-Akt, NF-κB and Bcl-2.Methods K562 cells were treated with cardamonin for 48 h, and the following tests were performed:(1) The cell morphology was observed by light microscopy.(2)IC50 of the K562 cells was dtermined by MTT test.(3) The apoptosis rate was detected by flow cytometry.(4) The expressions of Bcl-2 and Bax mRNA were detected a by RT-PCR.(5) The expressions of PTEN, p-Akt, NF-κB and Bcl-2 proteins were detected by Western blot.Results Obvious apoptosis was observed in the K562 cells after treated with cardamonin for 48 h.MTT assay indicated that the proliferation of K562 cells was obviously inhibited in a dose-and time-dependent manner. Comparing with the blank group, the early apoptosis rate and expression of Bax mRNA were significantly increased.At the same time, the expression of Bcl-2 mRNA was significantly decreased.All of them presented a dose-dependent manner. The expression of PTEN obviously increased with the increasing dose of cardamonin and the expressions of p-Akt, NF-κB and Bcl-2 were decreased.Conclusions Cardamonin promotes the apoptosis in K562 cells in a dose-dependent manner by increasing the expression of PTEN and decreasing the expressions of p-Akt, NF-κB, and Bcl-2.
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<p><b>BACKGROUND</b>Klebsiella pneumoniae carbapenemase (KPC)-producing Escherichia (E.) coli has been reported in China since 2008. However, there is no information about the molecular epidemiology of KPC-producing E. coli in China. In this study, we aimed to investigate the sequence type (ST) and characteristics of KPC-producing E. coli isolates in China.</p><p><b>METHODS</b>Three carbapenem-resistant isolates of E. coli (E1, E2, and E3) from one teaching hospital in Hangzhou covering a one year period were analyzed. Antibiotic susceptibility was determined by Etest. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were used for epidemiological analysis. The genetic structure around blaKPC, the major plasmid incompatibility typing, and the identification of β-lactamase gene types were performed by PCR and the positive products were subsequently sequenced. Plasmids were analyzed by transformation, restriction, and Southern blotting.</p><p><b>RESULTS</b>PFGE demonstrated that patterns of isolates E1 and E2 were clonally-related and designated as patterns A1 and A2; pattern of isolate E3 was different and designated as pattern B. MLST analysis showed that the three isolates displayed one common sequence type ST131. The identification of bla gene types by PCR and sequencing showed that blaKPC-2, blaCTX-M-14, and blaTEM-1 were detected in all three isolates. All three isolates carried a KPC-2-encoding plasmid of the IncN replicon. Plasmid analysis and hybridization experiments showed that the isolates were found simultaneously to carry two or four plasmids. The blaKPC-2 gene in E1 and E2 was located in a plasmid with size of ca. 50 kb. However, the blaKPC-2 gene in E3 was located in a plasmid with size of ca. 130 kb.</p><p><b>CONCLUSIONS</b>E. coli ST131 with KPC-2 β-lactamase has emerged in China, which enlarges the geographical area where the ST131 KPC-producing E. coli strains have diffused.</p>
Subject(s)
Bacterial Proteins , Genetics , China , Electrophoresis, Gel, Pulsed-Field , Escherichia coli , Genetics , Klebsiella pneumoniae , Multilocus Sequence Typing , beta-Lactamases , GeneticsABSTRACT
Objective To explore the application value of thromboelastography (TEG) in evaluation of coagulation status in late pregnancy women.Methods The results of TEG detection and coagulation function test were analyzed retrospectively in 84 late pregnancy women.The difference in detecting abnormal coagulation function was compared between the two methods.Results There were 10 late pregnancy women (11.9％) with abnormal coagulation function detected by TEG detection,and 26 (31.0％) with abnormal coagulation function detected by routine coagulation function test.The difference in detection rate of abnormal coagulation function was statistically significant between the two methods(x2 =4.53,P ＜ 0.05).Conclusion Hypercoagulable status in late pregnancy women detected by TEG detection was better than detected by routine coagulation function test.
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OBJECTIVE To investigate the prevalence of drug-resistant genes in seventeen strains of all-resistant Acinetobacter baumannii.METHODS Microdilute tests were performed to detect the susceptibility of 17 A.baumannii strains to 16 kinds of antimicrobial agents and antimicrobial-resistant genes were detected by PCR methods.RESULTS Seventeen A.baumannii strains showed all-drug resistance.Genes of TEM-1,OXA-23,OXA-27,gyrA and AmpC were detected in all 17 strains of A.baumannii.The positive rates of aacC31 and PER-1 genes were 11.8% and 52.9%,respectively.CONCLUSIONS A.baumannii with multi-resistant genes of our hospital carries TEM-1,OXA-23,OXA-27,gyrA,AmpC,aacC1 and PER-1.
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Objectives:To evaluate the relationship between the activity of cholesteryl ester transfer protein(CETP) and cardiocerebrovascular diseases. Methods:The CETP activity of 45 cases of coronary heart disease(CHD) and 26 cases of stroke were measured using the synthetic substrate of 14 C-radiolabeled discoidal bilayer particles as the cholesteryl ester donor and LDL as receptor. Results:The CEPT activity(x±s) of CHD and stroke patients were(17.6±5.4)% and(15.2±3.8)%,while the healthy controls were(12.7±2.0)%.The CEPT activities in CHD and stroke patients were significantly higher than that in healthy controls(P<0.01). Conclusions: CETP is related to the occurrence and development of atherosclerosis.