Objective To study the effect of pluripotency factor Nanog on the expression of the cell cy-cle related proteins,and then to explore its effect on the proliferative ability of HepG 2 cells.Methods TALENs gene editing tool was employed to induce mutation and downregulation expression of Nanog .T7 endonuclease 1 and genomic sequencing was used to analyze the mutation efficiency of Nanog .RT-PCR and western blot were used to determine the expression of mRNA and protein of Nanog ,respectively .Real-time cell based assay system was used to measure the proliferative ability of wild -type HepG2 cells and monoclonal HepG 2 cells with Nanog mutation.Results TALENs successfully induced mutation of Nanog gene .The targeting efficiency of mixed cells was analyzed by T7 endonuclease 1 approached 40%after two transfection with plasmid of Nanog -TALENs.Ad-ditionally,the Nanog mRNA expression level of monoclonal HepG 2 with Nanog mutation was downregulated by 3.4 times compared to the wild type HepG 2 cells,and the Nanog protein expression level was downregulated by 3.6 times.The cell cycle related proteins CyclinD1/D3,CyclinE1 and CDK2 expression were downregulated in monoclonal HepG2 with Nanog mutation in comparison to the wild type HepG 2 cells.Conclusion Nanog plays a role in influencing the proliferative ability of HepG 2 cells through modulating the expression of the cell cycle re-lated proteins CyclinD1/D3,CyclinE1 and CDK2.The downregulation expression of Nanog can inhibit the prolif-erative capacity of HepG 2 cells via the regulation of the cell cycle related proteins .

To elucidate the effect and effective mechanism of gonadotropin-releasing hormone analog (GnRHa) on growth axis and gonadal axis in puberty rats.

Objective To explore a kind of biomedical signal pattern (BSP) with a new method called as state representation methodology (SRM ) .Methods Based on the heart sound signals ,ECG signals ,breathing ,as the important research problem for BSP description ,with some comparisons on several traditional methods ,in which support vector machines (SVM ) and response sur-face methodology (RSM ) etc .,using frequency slice wavelet transform (FSWT ) method to extract the BSP signal dynamic damping characteristics ,thus ,this paper proposes a new idea of SRM analysis .In the case of heart sound signal analysis ,the general steps of SRM evaluation method is given .Results In 40 cases of normal heart sounds SRM model is set up ,with 80 cases of abnormal heart sounds are compared ,the obvious differences of the SRM state distributions of the two groups are found .Conclusion The combi-nation of SRM with FSWT can provide a novel approach for BSP analysis ,and provide powerful development tool for the analysis of BSP .

B-type ultrasound images have important applications in medical diagnosis. However, the widely spread intensity inhomogeneity, low-scale contrast, constructed defect, noise and blurred edges all make it difficult to implement automatic segmentation of lesion in the images. Based on region level set method, a subordinate degree region level set model was proposed, in which subordinate degree probability of each pixel was defined to reflect the pixel subjection grade to target and background respectively. Pixels were classified to either target or background by calculation of their subordinate degree probabilities, and edge contour was obtained by region level set iterations. In this paper, lesion segmentation is regarded as local segmentation of specific area, and the calculation is restrained to the local sphere abide by the contour, which greatly reduce the calculation complexity. Experiments on B-type ultrasound images showed improved results of the proposed method compared to those of some popular level set methods.
Algorithms ; Diagnostic Imaging ; Humans ; Models, Theoretical ; Probability ; Signal Processing, Computer-Assisted ; Ultrasonography

Objective To investigate the effect of recruitment maneuver(RM) on surfactant proteins in young piglets with acute lung injury and the possible mechanisms of lung recruitment after RM.Methods The piglet model of ALI was established by lipopolysaccharide intravenous injection,12 male piglets were randomly divided into two groups:conventional ventilation group(control group) and RM with low tidal volume group(RM group).After 8 hours of ventilation,mRNA expression of surfactant protein-A(SP-A),SP-B,SP-C,SP-D in the piglet lungs were determined by real time PCR and SP-A protein distribution was assessed by immunohistochemistry.Biochemical analyses of TP,total phospholipids(TPL),DSPC were conducted as well.SP-A levels in bronchoalveolar lavage fluid(BALF) and plasma were measured by enzymelinked immunosorbent assay(ELISA).Results As compared with control group,RM group had higher expression of SP-A,SP-B,SP-C and SP-D.SP-A average gray values of control group and RM group were 97.8±6.4 and 106.3±8.5,and there was significant difference(P＜0.01).RM group showed significant increase of TPL,DSPC and DSPC/TP.The concentration of SP-A in BALF was higher in RM group than that of the control group,however,SP-A plasma level was lower in RM group than that of the control group.Conclusion RM can increase suffactant protein expression in ALI animals,alleviate surfactant protein dysfunction and regulate the concentration of SP-A,which may improve alveolar recruitment following the RM and alleviate ventilator-induced lung injury.

OBJECTIVE To investigate the pathogenic distribution and drug resistance of nosocomial infection occurred in pediatric patients with hematologic malignancy and therefore provide the information in rational administration of antibiotics to pediatric patients with hematologic malignancy complicated with nosocomial infection.METHODS Flora cultivation and isolation were operated with the routine methods and drug-sensitivity was determined by Kirby-Bauer method.RESULTS Totally 116 strains of pathogenic bacteria were isolated,in which included 78 strains of G-bacteria and 38 strains of G+ bacteria.Fungi were also very common.In this study,both G-and G+ bacteria were resistant to antimicrobial agents tested.CONCLUSIONS The bacterial spectrum and their drug-resistance characteristics in pediatric patients with hematologic malignancy are quite different to that encountered in pediatric patients with other systemic diseases.Most strains present high resistance to antibiotics,so our administration of antibiotics for nosocomial infections should be directed and carry out according to the susceptibility tests in different area and different periods.

Objective:To explore the role of 6-CYANO-2,3-DIHYDROXY-7-NITROQUIN OXALINE (CNQX) in different types of synapse secretion.Methods:The spontaneous mEPSCs and eEPSCs at different extracellular concentrations of CNQX in cultured cortical or hippocampal neurons were recorded respectively.Results:The half inhibitory concentration (IC50) of CNQX in evoked neurotransmitter release was significantly higher than that of spontaneous release,indicating that the spontaneous neurotransmitter release was more sensitive to CNQX.No apparent difference was observed between cortical and hippocampal cells,suggesting that the blocking effect of CNQX was similar in different brain regions.Conclusion:CNQX might have differential regulating mechanisms between excitatory spontaneous and evoked neurotransmitter release,but without brain regions specificity.

AlM: To explore the expression and significance of cysteine- rich 61 ( CCN1/Cyr61 ) in oxygen - induced retinal neovascularization ( RNV) of mice and study the inhibition effect of CCN1 specific siRNA on RNV.
METHODS:Two hundred healthy C57BL/6J mice were chosen and randomly divided into control group, hyperxia group, hyperxia control group and CCN1 treated group, with 50 mice in each group. The hyperxia control group was treated with vector plasmids by intravitreal injection. The CCN1 treated group received CCN1 siRNA recombinant plasmids by intravitreal injection. Adenosine diphosphate-ase ( ADPase) stained retina flat-mounts was performed to assess the retinal vascular profiles, HE staining was applied to count the number of vascular endothelial cell nuclei breaking through the internal limiting membrane, protein and mRNA level expression of CCN1 were measured by immunohistochemistry, Western blot and RT-PCR.
RESULTS: There were large nonperfusion area and a large number of vascular endothelial cell nuclei breaking through the internal limiting membrane ( 25. 25 ± 1. 26;23. 12 ± 1. 16 ) in the hyperxia group and the hyperxia control group. Regions of nonperfusion and vascular endothelial cell nuclei (8. 47±1. 15) were decreased in the CCN1 treated group compared to the hyperxia group and the hyperxia control group. Compared with the control group, there were high protein and mRNA expression of CCN1 in the hyperxia group and the hyperxia control group. The expression of CCN1 protein and mRNA were decreased in the CCN1 treated group compared with the hyperxia group and hyperxia control group (all P<0. 05).
CONCLUSlON: The abnormal expression of CCN1 has close relation with RNV. The development of RNV can be markedly inhibited by RNA interference targeting CCN1, which, we believe, will provide new molecular targets and a rationale for clinical developing new strategy for ROP therapy.

Mild cognitive impairment (MCI) is a transitional stage between normal aging and dementia. The main characteristic of the patients with MCI is the impairment of episodic memory in which hippocampus plays an important role. Therefore, the detection of structural and functional changes of hippocampus will be the key to early diagnosis of MCI. This paper presents a brief overview of recent study about hippocampal magnetic resonance imaging of MIC.

Objective By detecting the variation of long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) DNA methylation in preeclampsia-like mouse models generated by different ways, to explore the roles of multifactor and multiple pathways in preeclampsia pathogenesis on molecular basis. Methods Established preeclampsia-like mouse models in different ways and divided into groups as follows: (1) Nw-nitro-L-arginine-methyl ester (L-NAME) group: wild-type pregnant mouse received subcutaneous injection of L-NAME;(2) lipopolysaccharide (LPS) group:wild-type pregnant mouse received intraperitoneal injection of LPS; (3) apolipoprotein C-Ⅲ (ApoC3) group: ApoC3 transgenic pregnant mouse with dysregulated lipid metabolism received subcutaneous injection of L-NAME;(4)β2 glycoprotein I (β-2GPI) group:wild-type pregnant mouse received subcutaneous injection ofβ-2GPI. According to the first injection time (on day 3, 11, 16 respectively), the L-NAME, LPS and ApoC3 groups were further subdivided into:pre-implantation (PI) experimental stage, early gestation (EG) experimental stage, and late gestation (LG) experimental stage.β-2GPI group was only injected before implantation. LCHAD gene methylation levels in placental were detected in different experimental stage. Normal saline control groups were set within wild-type and ApoC3 transgenic pregnant mice simultaneously. Results (1) CG sites in LCHAD DNA:45 CG sites were detected in the range of 728 bp before LCHAD gene transcription start site, the 5, 12, 13, 14, 15, 16, 19, 24, 25, 27, 28, 29, 30, 31, 32, 34, 35, 43 CG sites were complex sites which contained two or more CG sequences, others were single site which contained one CG sequence. The 3, 5, 6, 11, 13, 14, 18, 28 sites in L-NAME, LPS, ApoC3 and β-2GPI groups showed different high levels of methylation; the 16, 25, 31, 42, 44 sites showed different low levels of methylation; other 32 sites were unmethylated. (2) Comparison of LCHAD gene methylation between different groups:the methylation levels of LCAHD gene at 3, 11, 13, 14, 18 sites in L-NAME, LPS, ApoC3 andβ-2GPI groups were significantly higher than those in the normal saline control group (P<0.05); and the methylation levels of 42, 44 sites in these groups were significantly lower than those in the normal saline control group (P<0.05). (3) Methylation of LCHAD gene at the same site between different experimental stages: ① The 3, 11, 18 sites of EG experimental stage was significantly lower than PI and LG experimental stage in L-NAME group (P<0.05);the 3, 11, 18 sites of PI experimental stage was significantly lower than EG and LG experimental stage in LPS group (P<0.05);these sites of PI experimental stage was significantly higher than EG and LG experimental stages in ApoC3 group (P<0.05).②The methylation of site 5 in L-NAME and LPS groups were significantly higher than that of the normal saline control group (P<0.05), and the LG experimental stages were significantly higher than other stages, but in ApoC3 group , only PI and EG stages were significantly higher than the normal saline control group (P<0.05).③At site 6 in L-NAME group which showed high methylation level was significantly higher than the same site in other groups which showed low methylation level (P<0.05).④At 13, 14 sites, earlier preeclampsia onset caused a lower methylation level in L-NAME group, but PI experimental stage was significantly higher than EG and LG experimental stages in LPS group (P<0.05), EG experimental stage was significantly higher than PI and LG experimental stages in ApoC3 group (P<0.05). ⑤ At site 28, earlier preeclampsia onset caused a higher methylation level in L-NAME group, but PI experimental stage was significantly lower than EG and LG experimental stages in LPS group (P<0.05), EG experimental stage was significantly higher than PI and LG experimental stages in ApoC3 group (P<0.05).⑥The 16, 25, 31 sites in ApoC3 group were significantly higher than other groups (P<0.05). ⑦ At site 42 in β-2GPI group was unmethylated, but it in other groups showed low methylation level, the methylation level of site 42 inβ-2GPI group was significantly lower than that in other groups (P<0.05). Conclusions The methylation of 6 and 42 CG sites may be related to LCHAD gene expression in placenta of L-NAME and β-2GPI induced preeclampsia-like models respectively;LCHAD gene expression and DNA methylation may not have obviouscorrelation in LPS and ApoC3 induced preeclampsia-like models. Differences exist in LCHAD DNA methylation in preeclampsia-like models generated by different ways, revealed a molecular basis to expand our understanding of the multi-factorial pathogenesis of preeclampsia.