Objective To investigate the role of HMGB 1 in epithelial-mesenchymal transition .Methods Specific siRNA of HMGB1 was designed and synthesized .Cultured typeⅡalveolar epithelial cell line-A549 cells were divided into 4 groups:1)control group, 2)model group induced by TGF-β1, 3)HMGB1 RNAi group, 4)RNAi negative con-trol group .Cellular morphology changes were observed by phase-contrast microscope .HMGB1 andα-SMA expression in A549 cells was detected by RT-PCR and Western blotting respectively .Results mRNA and protein expression of HMGB1 andα-SMA in TGF-β1 group increased significantly than that in control group (P<0.01).HMGB1,α-SMA mRNA and protein expression in siRNA-treated cells decreased significantly as compared with that in TGF -β1 group (P<0.01).Conclusions HMGB1 may be involved in the TGF-β1 induced EMT.

Hepatocellular carcinoma ( HCC) is a prevalent primary liver cancer that is driven by cumulative chan-ges in the hepatocyte genome which were influenced by the liver microenvironment .The tumor microenvironment is a dynamic system, including cancer cells, stroma, the extracellular matrix.tissue hypoxia, diet, gastrointestinal tract ( GIT) microflora and circulating microvesicles , The liver microenvironment plays a pivotal role in HCC tu-merigenesis , progression and therapeutic efficienry .The study of the HCC microenvironment will provide new in-sights into the mechanism of tumourigenesis and potential novel targets in the treatment of HCC .

Radiation-induced cognitive impairment is hypothesized to occur because of dynamic interactions be -tween multiple cell types, including astrocytes, endothelial cells, microglia, neurons, and oligodendrocytes.Cur-rent researche indicates that radiation-induced changes include the decrease in hippocampus neurogenesis , altera-tions of neuronal functions , particularly synaptic plasticity , as well as the elevation of neuroinflammatory cytokines .

Diabetic cognitive dysfunction is one of the important complications of diabetes mellitus .Insulin plays an important protective role in cognitive function through MAPK and PI 3-K/Akt signaling pathway .Insulin resistance may give rise to excessive tau protein phosphorylation , formation of neurofibrillary tangles inhibits insulin degrading enzyme , reduces the degradation of amyloid β;enhances oxidative stress , further damages the integrity of the neu-ronal structure and function and eventually leads to cognitive dysfunction .

Objective To study the strategy of sequential invasive-noninvasive mechanical ventilation in chronic ob-structive pulmonary disease ( COPD ) and specially to explore the role of mechanical ventilation in patients with weaning difficulty .Methods Forty-five cases withe weaning difficulty during January 2009 ~December 2013 from Huludao central hospital emergency section guardianship room were recruited .Compare the rate of re-tracheal intu-bation rate and withdrawal machine success rate in two groups ( intervention group , n=21 , control group , n=24 ) . Results In the intervention group after weaning the 24 h pH, PaCO2 was significantly better than that of the control group ( P<0.01 ) ,in the endotracheal intubation intervention group ,48 h re-tracheal intubation rate was 9.5%, less than the control group ( 41.7%) (P<0.05), The relative risk of intervention group was 0.147 (95%CI, 0.028~0.781 ) .Intervention weaning success rate is significantly higher than that of the control group (85.7%vs 50.0%,P<0.05 ) .Conclusions Sequential invasive-noninvasive mechanical ventilation strategy reduces 48 h in-tubation rate and improve the success rate of weaning .

Objective To explore the impact of silencing PLCε1 gene on proliferation and cell cycle of esophageal carci-noma Eca109 cells.Methods Three plasmid expression vectors (PLCε11, PLCε12 and PLCε13) were constructed to si-lence PLCε1 gene.A negative control plasmid expression vector (HK) was constructed at the same time to serve as a control .The plasmid expression vectors were transfected into esophageal carcinoma Eca 109 cells by cations liposome . The plasmid expression vector with the best interference effect ( PLCε12 ) was chosen .The study included Eca 109 group , HK group and PLCε12 group .Cell viability of Eca 109 cells was evaluated by MTT assay .The cell cycles were detected by FCM .The mRNA expression of P16 and CyclinD1 gene was measured by RT-PCR.Results The cell vi-abilitys of Eca109 cells in PLCε12 group were 80.73%and 75.88%at 48 and 72 h after transfection , which were significantly lower than that of Eca 109 cells in HK group (P<0.001).The percentage of S phase Eca109 cells in PLCε12 group was lower than that of Eca 109 cells in HK group ( P <0.01 ) , the cell cycle of PLCε12 group Eca109 cells was arrested in G0/G1 phase.The P16 gene mRNA expression of PLCε12 group Eca109 cells was higher than that of HK group Eca 109 cells ( P<0.01 ) .Conclusions Silencing PLCε1 gene may up-regulate P16 gene mRNA expression and then arrest the cell cycle at G 0/G1 phase and so inhibit proliferation of Eca 109 cells.

Objective To determine the effects of EGCG on lipopolysaccharide ( LPS)-induced neuroinflamma-tion and investigate the role of neuroprotection mediated by EGCG .Methods Primary cultures of rat gliacyte were used as an in vitro model to examine the effects of EGCG on LPS-induced neuronal damage .The intracellu-lar Glu andγ-GABA were quantified via HPLC .Then the protein level of TNF-α,IL-1βand IL-8 was determined by ELISA and Western blot assay .Results Compared with the control group , LPS apparently induced the pro-duction of intracellular ROS ( P<0.05 ) and released the TNF-α, IL-1βand IL-8 in the primary cultures super-natant (P<0.05).Compared with the LPS group,EGCG significantly attenuated the release of TNF-α, IL-1βand IL-8 ( P<0.05 ) and the level of iNOS protein ( P<0.05 ) .LPS apparently induced the production of intra-cellular ROS( P<0.05 ) and released the TNF-α, IL-1βand IL-8 in the primary cultures supernatant ( P <0.05 ) .EGCG significantly attenuated the release of TNF-α, IL-1βand IL-8 ( P<0.05 ) and the level of iNOS protein(P<0.05), and rugulated the concentration of Glu/γ-GABA(P<0.05).Conclusions EGCG is effective in protecting hosts against LPS-induced neuroinflammation through anti-inflammatory effects and regulating extracel-lular Amino acid levels .

Objective To explore whether quercetin-loaded PEG-PE micelles(M-Q) can synergize the growth-in-hibitory activity of adriamycin prepared ( ADR) by reversing the drug resistance of MCF-7 ADRr breast cancer cells in vitro.Methods M-Q was prepared by adding saline to lipid film containing quercetin and PEG-PE.The size of M-Q was characterized by dynamic light scattering ( DLS) .The inhibition of MCF-7 ADRr cells was evaluated by MTS assay after incubation with M-Q and ADR.Results The incorporation efficiency of quercetin by the micelles was above 74%.The average size of M-Q was 11.11 nm.Compared with the quercetin dissolved in ethanol , M-Q more effectively reversed the drug resistance of MCF-7 ADRr cells in vitro.Conclusions PEG-PE micelles may potentially deliver quercetin to cancer cells for reversal of drug resistance .

Objective To construct eukaryotic vectors expressing short hairpin RNAs ( shRNAs ) targeting at the CIP2 A gene and to explore its effects on gastric cell line BGC-823 .Methods Four oligonucleotides targeting the CIP2A gene were synthesized and cloned into the eukaryotic expression plasmid pGPU 6.The recombinant plas-mids, pGPU6/GFP/Neo-CIP2A-shRNA-1, 2, 3 and 4, were introduced into BGC-823 cells by lipofectamine-me-diated transfection and the infection rate was observed by fluorescence microscope .The gene silencing efficiency was measured by real-time PCR and Western blot .The effects on proliferation of BGC-823 cells were detected by CCK-8 .Results DNA sequencing and enzyme digestion analysis confirmed the identity of the four recombinant shRNA expression vectors .Immunofluorescsence demonstrated that transfection efficiency was above 70%.Trans-fection of shRNA-1, 2, 3 and 4, significantly knocked down the expression of CIP 2A mRNA and protein at 24, 48 and 72 h after transfection .Compared with the 2 , 3 and 4 , shRNA-1 had the more strong inhibitory effect on the expression of CIP2A mRNA and protein.The CCK-8 assay showed that the anti-proliferation effect on BGC-823 cells was significant ( P<0.05 ) .Conclusions The recombinant vector may effectively inhibit the expression of CIP2A in BGC-823 cells and depress the proliferation of BGC-823 cells.

Objective To investigate the effect of notch signaling pathway on drug resistance and invasiveness of bladder cancer .Methods We observed the changes of growth and morphology of bladder cancer T 24 , 5637 and J82 cells which treated for 48 hours using γ-secretase inhibitor by inverted microscope .The mRNA and protein lev-els of the EMT molecular markers , including E-cadherin , N-cadherin , vimentin and Alpha-smooth muscle actin were examined by RT-PCR and Western blot in bladder cancer cells;Detected the changes of drug resistance and invasion respectively by MTT and Transwell in bladder cancer cells .Results After completely blocking the Notch signaling pathway , the inverted microscope showed that bladder cancer cells became smaller and more disperse ;RT-PCR and Western blot showed the mRNA and protein levels of E-cadherin were up-regulated ( P<0.05 ) , contrast , N-cadherin , vimentin and Alpha-smooth muscle actin were down-regulated ( P<0.05 ); The prolifera-tion of bladder cancer cells were significantly inhibited by MTT test;The number of through microporous membrane cells significantly decreased ( P<0.05 ) shown by Transwell test .Conclusions The Notch signaling pathway is completely blocked that nhibites proliferation and EMT of bladder cancer cells , reduces drug resistance and inva-sion in bladder cancer cells .It suggests that drug resistance and invasiveness of bladder cancer can be changed through EMT which is regulated through notch signaling pathway .