Objective To analyze neural activity of in rat prefrontal cortex with the use of nonnegative matrix factorization with sparseness constrains (NMFs) as a methodology and to study how to express neural ensemble with higher precision during working memory task.Methods Experiment data were obtained from neural population activity in the period 5 s before and after the working memory event.From the zero point,the neuronal firing times were binned in windows of 200 ms with 50 ms overlapping.The normalized neuronal bin-count matrix is decomposed by NMFs into mixing matrix and source component matrix with sparseness constraints.Meaningful components were extracted to reconstruct the input by an inverse of NMFs transform.Results By analyzing the ten groups of data from 2 rats,with the numbers of the sparse sources of 10 and 15 respectively,explicit neural ensembles with the feature components were obtained in the sparse reconstructed activity.Comparing to rate coding,the spatiotemporal location of neural ensemble was more precisely detected.Conclusion The working memory information is encoded with neural ensemble activity.NMFs could find the sparse firing pattern robustly in neuron population activity.NMFs removes much redundancy and demonstrate the possibility to express neural ensemble with higher precision compared with rate coding,which would be helpful to infer correlations between cortical firing pattern and working memory event.

Objective To investigate how the independent components(ICs)energies of multichannel local field potentials(LFPs) code event base on the analysis of ICA of the cortical LFPs of rats. Methods Taking the event point as the zero point, 15-channel LFPs between the span of ±500ms recorded from the prefrontal cortex of rats were decomposed into 15 ICs. The energies of the ICs were computed in a 50-ms window. By sliding the window with step of 25 ms, a dynamic distribution mapping of the 15 ICs' energies was established. ICs with distinctly increased energies during the span of ±200 ms, which indicating that these ICs energies coded event,were selected as the targets. The corresponding channels of these ICs were determined consequently via the inverse transformation of ICA. Results Considering each trail of the repetitious analysis for the same segment of data, the spatial localization of the dominate function region(s) turned out to be relatively stable in spite of the uncertainty of the number and sequence of the target IC(s) due to the ambiguities of the decomposition of ICA.Meanwhile, the analysis results of a series of data segments showed satisfactory correspondence between data segments and dominate function regions. Conclusion The ICs' energies of multichannel LFPs are able to code events in working memories; It is valid for ICA to identify the coding patterns of multichannel LFPs to events; ICA is capable to localize the dominate function regions of event coding with satisfactory robustness.

Objective To investigate the character and location of a novel gene R049 and its expressed protein in uropathogenic Escherichia coli(UPEC) strain 132 isolated in China. MethodsThe chromosome library of UPEC132 was constructed by a shotgun strategy and the sequence analysis was carried out by a high-throughput pyrophosphate sequencing. Sequence reads were assembled with the Newbler program.The characters of R049-associated specific fragment were analyzed using the bioinformatics methods. Outer and inner membrane proteins of UPEC132 were extracted and then detected by SDS-PAGE and Western blot analysis together with the whole-cell lysates. ResultsThe 169 022 bp contig containing gene R049 was obtained and its sequence was very similar to the chromosome associated sequence of UPEC strain 536. It showed that a 20 773 bp fragment including R049 replaced the pathogenicity island PAI Ⅲ536 of UPEC536 in above 169 022 bp contig. The fragment had a lower GC content (46.97％) and 16 bp direct repeats in two ends. Significantly it also was adjacented to thrW tRNA, insertion element and genes coding integrase. Thus the 20 773 bp fragment was named R049 genome island(R049-GI). There were 25 ORFs in R049-GI, and gene R049 was located in the thirteenth ORF. The results of SDS-PAGE and Western blot revealed gene R049 encoded an outer membrane protein in the size of 47.0× 103. ConclusionGene R049, encoding an outer membrane protein, was a component part of the genome island in UPEC 132 chromosome acquired by horizontal gene transfer.

Objective Toinvestigatetheentropyoflocalfieldpotentials(LFPs)recordedinratmedialprefrontal cortex during a Y-maze working memory (WM) task, to provide computing support for neural coding mechanism.Methods Sixteen-channel LFPs were recorded from SD rats while they performed a Y-maze WM task.The data came from 4 rats, 20 trials (10 correct trials and 10 incorrect trials) per rat provided by laboratory of neurobiology in medicine,Tianjin Medical University.Original LFPs were preprocessed to remove 50 Hz power line noise and baseline drift.Multi-taper Fourier transform was applied to calculate spatial distributions of LFPs and band pass filter were used to extract characteristic signal.The entroy coding of 16 channel LFPs was as follows: the physiological window was set to be 500 ms, the step length of physiological window was set to be 125 ms, windows were added to LFPs data, and then LFPs entropy of each sliding window was computed and averaged to get the trend of multichannel entropy values duringthe WM task.Results The power of θ band (4-12 Hz) in LFPs increased.The averaged entropy value ofmultichannel θ band LFPs in correct trials was 0.939±-0.020, which were larger than those in the resting state, 0.795±0.031 (P＜0.05).Those during wrong WM task had no significant difference, which didn't encode the WM task.Conclusions The principal frequency band related to WM is the θ band and LFPs entropy encodes the WM effectively.

Objective Infrared spectroscopy was used to study on both raw material and different degree water processing varieties extract of Radix Aconiti kusnezoffii,to observe the changes of main toxic components in processing,and thus to improve the quality of Radix Aconiti kusnezoffii processed products.Methods Fu Liye transform infrared spectroscopy was adopted to study the infrared spectrum characteristic of raw materials and different degree water processing varieties of.Radix Aconiti kusnezoffii.Results ①Aconitine and hypaconitine infrared spectrum showed that 1717 cm-1,1727 cm-1 and 1711 cm-1 is the C=O stretching vibration peak.It is a diester alkaloids characteristic peaks; ② although absorption peak of all vesicular samples had a certain change,it still existed diester alkaloids absorption peak,indicating the incomplete hydrolysis; ③ boiled aconite processing methods demonstrated diester alkaloids absorption peak shift in the water sample.Conclusion Diester alkaloids in Radix Aconiti kusnezoffii shows a positive relation with its time soaked in water.

Aim To investigate the effect of Jinlida on cholesterol-related genes in skeletal muscle in fat-in-duced insulin resistance ApoE-/ - mice. Methods Ten male C57 BL/6 J mice were selected as normal group ( NF );50 male ApoE-/ - mice with a high-fat feeding after 16 weeks ( HF) were divided into model group, rosiglitazone ( LGLT ) , Jinlida low dose group ( JLDL, 0. 95 g · kg-1 · d-1 ) , Jinlida medium dose group ( JLDM, 1. 9 g·kg-1 ·d-1 ) , Jinlida high dose group (JLDH, 3. 8 g·kg-1·d-1), which were per-formed intragastric administration for 8 weeks. Oil red O staining of mouse skeletal muscle was used for fat ac-cumulation. Insulin receptor ( INSR) , insulin receptor body substrate-1 ( IRS-1 ) , low-density lipoprotein re-ceptor ( LDLR ) , cholesterol sensor ( SCAP ) mRNA and protein expression in mouse skeletal muscle were measured by quantitative reverse transcription PCR ( RT-PCR ) and Western blot. Results Compared with NF group, fasting blood glucose ( FBG) , choles-terol ( TC ) , triglyceride ( TG ) and low density lipo-protein cholesterol ( LDL-C ) of HF mice were signifi-cantly elevated, while high-density lipoprotein ( HDL-C ) significantly decreased ( P < 0. 05 ) . Compared with HF group, Jinlida group could reduce to varying degrees FBG, TC, TG and LDL-C in mice, and in-crease HDL-C ( P <0. 05 ) . Jinlida could downgrade fasting serum insulin ( FINS ) level, and improve the insulin sensitive index ( ISI ) ( P < 0. 05 ) . Jinlida could obviously improve skeletal muscle fat accumula-tion of mice. Compared with NF group, skeletal mus-cle INSR, IRS-1, LDLR mRNA and protein levels of HF group were significantly decreased ( P <0. 05 ) , while SCAP mRNA and protein level increased signifi-cantly (P<0. 05). Compared with HF group, Jinlida could increase to varying degrees INSR, IRS-1, LDLR mRNA and protein levels ( P < 0. 05 ) , and lower SCAP mRNA and protein levels ( P<0. 05 ) . Conclu-sion Jinlida can alleviate fat-induced insulin resist-ance in ApoE-/ - mice through regulation of cholester-ol-related gene expression.

OBJECTIVE To explore the effect of Lianhuaqingwen capsules ( LHQW ) on junction protein expression in mouse lung tissue of lipopolysaccharide (LPS)-induced acute lung injury ( ALl). METHODS 120 male mice were randomly divided into six groups: normal control, model, model+dexa-methasone 5 mg.kg-1 , model +LHQW 2, 4 and 8 g.kg-1 groups. Dexamethasone and LHQW were administered orally, once daily, for 7 d. 24 h after the last administration, LPS solution was instilled into the tracheas of mice except the normal control group to prepare the mouse model of ALl. 24 h after the establishment of the ALl model, the mice were sacrificed and the pathological changes in the mouse lung tissue were observed by optical microscopy and ultrastructure of alveolar epithelium was observed by transmission electron microscopy. The cell percentage of positive expression of tumor necrosis factor-α(TNF-α) in the peripheral blood T lymphocytes was detected by flow cytometry. The expressions of con-nexin 43 ( Cx43), occludin and zonula occludens protein-1 ( ZO-1) in lung tissues were detected by immunohistochemistry. RESULTS Under the light microscope, the mouse lung of model group showed a large amount of inflammatory cell infiltration and alveolar wall thickening. Compared with model group, inflammatory cell infiltration was reduced in model+dexamethasone, model+LHQW 2,4 and 8 mg.kg-1 groups. Under the electron microscope, the mouse alveolar epithelial cells of model group showed injury. Compared with model group, the damage was reduced in model+dexamethasone, and model+LHQW 2, 4 and 8 mg.kg-1 groups. The cell percentage of TNF-α positive expression in peripheral blood T lympho-cytes in normal control, model, model+dexamethasone, model+LHQW 2,4 and 8 mg.kg-1 groups was (3.6±0.9)%, (6.4±0.8)%, (2.8±0.7)%, (4.7±1.6)%, (4.0±1.5)% and (3.6±1.2)%, respectively. The percentage in model group was obviously higher than that in normal control group( P<0.01), but was lower in the four drug treatment groups than in model group(P<0.05, P<0.01). The expression of Cx43, occludin and ZO-1 in lung tissue of model group was lower than that of normal control group(P<0.01), but higher in model+dexamethasone, model + LHQW 4 and 8 mg.kg-1 groups than in model group(P<0.05). CONCLUSION LHQW may alleviate ALl induced by LPS and play a protective role by inhibiting inflammatory cell infiltration and improving protein connection expression in alveolar epithelial cells and pulmonary vascular endothelial cells.

Objective To compare the differences of lung pathological changes of acute lung injury in mice in-duced by lipopolysaccharide ( LPS) and graphite particles, and to explore the possible mechanisms of acute lung injury in-duced by fine particles of different origins.Methods 140 male specific-pathogen-free Kunming mice weighing 18-20 g were randomly divided into 7 experimental groups, in addition to the normal control group.The experimental groups were treated by intratracheal instillation of LPS solution or graphite powder suspension in different doses, respectively, to induce acute lung injury in the mice.The mortality of the mice was observed, and pathological changes of the lung tissues were ex-amined by light and transmission electron microscopy.Western blot was used to detect the protein expression of neutrophil elastase ( NE) in lung tissues , and real-time quantitative PCR was used to detect mRNA expression of monocyte chemotac-tic protein-1 ( MCP-1) in the lung tissue .Results Compared with the normal control group, some pathological changes were observed in the lung tissues of the groups L ( LPS) and G ( graphite) .There were numerous macrophages in the lung tissues in the group G mice, and exudate, mainly neutrophils, in the lung tissues of the group L.The NE protein expres-sion in the lung tissue was significantly higher than that of the normal control group ( P<0.05) , and there was also a sig-nificant difference between the groups L and G (P<0.05).The MCP-1 mRNA expression in lung tissues was higher in the control group (P<0.01), and there was also a significant difference between the groups L and G (P<0.01).Conclu-sions Diverse types of particulate matters induce different pathological changes in the lungs, therefore the mechanism may also be different in the inflammatory responses.It means that the lung injuries caused by fine particles of mixed composition may have complex mechanisms.

Objective The aim of the study was to investigate the early failure of newly created arteriovenous fistula(AVF) in patients on maintenance hemodialysis and the factors responsible for the failure. Methods The clinical data were retrospectively reviewed, preoperative examination and laboratory biochemical indicators of 88 patients with newly created AVF for maintenance hemodialysis in our hospital through Hospital Management Information System and telephone follow-ups. Binary Logistic regression was used to analyze the protective factors for early failure. Results In 88 patients, early failure of the AVF was found in 15 patients. Twenty-three factors, including gender, were involved in statistical analysis. There were statistical differences between the two groups in hypertension (χ2=7.689, P=0.006) and whether they had early referral to nephrologists (χ2=5.334, P=0.021). Further regression analysis showed hypertension ( OR=0.192, 95% CI=0.0538-0.692, P=0.012) was protective factor and without early referral ( OR=3.651, 95% CI=1.068-18.302, P=0.039) was the risk factor of early failure. Conclusion This study shows that no early referral and combined hypertension is an important factor affecting the early failure, emphasizing the clinical work, for the diagnosis of patients with chronic kidney disease, early nephrological referral should be established, and blood pressure monitoring should be done to help reduce the incidence of complications.

Objective To construct protective immunity to Mycobncterium tuberculosis latent infection, a novel fusion protein consisting of HspX, the 190 to 198 peptide of Mpt64 and Ag85B, which were confirmed to be the effective protective antigens mainly expressed in the dormancy and exponential phase of growth, was constructed and its immunogenicity was investigated. Methods Ag85B and Mpt64190-198-HspX sequences were amplified by PCR and cloned into plasmids pET-28a. The fusion protein, Ag85BMpt64190-198-HspX (AMH) was expressed in E. coli BL21 and purified with Ni-NTA resins. C57BL/6 mice were immunized three times at 2-week intervals subcutaneously with AMH formulated with the adjuvant composed of dimethyl-dioctyldecyl ammonium bromide (DDA) and BCG polysaccharide nucleic acid (BCGPSN). Humoral and cell-mediated immunity responses were analyzed at five weeks after the last injection. Results AMH was expressed stably in E. coli and could be purified well by Ni-NTA affinity chromatography. C57BL/6 mice immunized with AMH subunit vaccine generated specific cellular and humoral immunologic response to the stimulation of Ag85B, Mpt64190-198 and HspX. Conclusion It suggested that AMH was a promising candidate antigen of tuberculosis subunit vaccine.