During the process of pathogens penetrating the plant cell, pathogens often secret some chemicals into plant cells, at the same time, they also produce mechanical signal by physical pressure on the plant cell. Here the pressure is used as the stress signal, to study its effect on phytoalexin accumulation and the induction of plant resistance in cucumber seedling. It is found that stress can induce the resistance in cucumber seedling significantly. When breaking the plant cell wall and plasma membrane adhesion by RGD peptides, the resistance induction is almost eliminated. Results from TLC and HPLC showed that stress stimulation could increase phytoalexin accumulation in cucumber seedling. This suggests that the accumulation of phytoalexin is one possible reason of the increased resistance after stress stimulation. When the adhesion between plant cell wall and plasma membrane was block by RGD, there is only small amount of phytoalexin accumulation compared with the control, suggesting that the stress induced phytoalexin accumulation and resistance is relying on the adhesion of plant cell wall and plasma membrane.

ABSTRACT:Objective To investigate changes in the neutrophils in rats with D-galactosamine (D-GalN)-induced acute liver failure (ALF)and to explore the therapeutic effect of interventions treatment of neutrophils on ALF.Methods Liver function,the expressions of inflammatory cytokines TNF-αand IL-1β,and the changes of neutrophils in the peripheral blood and the liver were observed in rats with D-GalN (intraperitoneal injection)-induced ALF.SD rats were randomly divided into three groups when treated with intervention of neutrophils:control group,ALF group (intraperitoneal injection of D-GalN),and treatment group (intravenous injection of anti-PMN serum via tail vein 24 h before modeling).Biochemical analysis was used to detect serum ALT,AST, TBIL and blood ammonia.Hematology analyzer was applied to analyze the number and percentage of peripheral blood neutrophils.The number of neutrophils in the liver was evaluated by immunohistochemistry.Liver RT-PCR was adopted to detect the mRNA expression of inflammatory cytokines TNF-αand IL-1β.Results We found that 6 h after D-GalN injection,serum ALT,AST,TBIL and blood ammonia in ALF rats were significantly increased (P <0.05).The mRNA expression levels of inflammatory cytokines TNF-αand IL-1βin the liver reached the peak at 6 h after modeling (P <0.001),and it was still notably higher at 24 h than before modeling (P <0.001 ).The number and percentage of peripheral blood neutrophils and the number of neutrophils in the liver were all markedly increased 12 h after modeling (P <0.001 ),and the increase continued at least until 24 h (P <0.001 ).24 h after intravenous injection of anti-PMN serum via tail vein,ALF rats had a distinct decrease in the number of peripheral blood neutrophils and neutrophils in the liver 24 h after modeling (P <0.001).Meanwhile,serum ALT,AST,TBIL and blood ammonia were all greatly decreased compared with those in ALF group (P <0.05);a significant reduction of hepatocyte apoptosis was observed.Also,the expressions of TNF-α and IL-1β in the liver were remarkably decreased after treatment (P <0.05).Conclusion Neutrophils accumulated in peripheral blood and liver of rats with D-GalN-induced ALF.The treatment of anti-PMN serum may have a therapeutic effect on liver function and immune microenvironment in ALF rats.

BACKGROUND:Cel co-culture can maximize the simulation ofin vivomicroenvironment. Cel scratch test and interleukin-1β can destroy the balance between matrix metaloproteinases (MMPs) and matrix metaloproteinase inhibitors (TIMPs), resulting in extracelular matrix degradation of the articular cartilage, functional disorders of chondrocytes and articular cartilage degeneration. OBJECTIVE:To study the effect of interleukin-1β on migration, MMP and TIMP expression of chondrocytes co-cultured with osteoblast supernatantin vitro. METHODS:There were three groups: chondrocyte monoculture group, osteoblast+chondrocyte group (co-culture group), osteoblast+chondrocyte+interleukin-1β group (interleukin-1β group). Cel scratch test was conducted to observe the migration of chondrocytes within 24 hours. Semi-quantitative PCR test was used to detect the changes in expressions of MMP-1, MMP-2, MMP-3, MMP-9, TIMP-1, TIMP-2, TIMP-3, TIMP-9 in chondrocytes within 24 hours. RESULTS AND CONCLUSION:Compared with the monoculture group, cel migration rate of the other two groups were increased significantly (P< 0.01). Compared with the monoculture group, the gene expressions of MMP-1, MMP-2, MMP-3 and MMP-9 were increased significantly in the coculture group (P < 0. 05); the gene expressions of MMP-1, MMP-3, MMP-9 were increased significantly in the interleukin-1β group (P< 0. 01). Compared with monoculture group, the gene expression of TIMP-1 was increased significantly (P < 0. 01), but the gene expressions of TIMP-3 and TIMP-4 were declined significantly (P < 0. 05) in the other two groups. These findings indicate that co-culture of chondrocytes with osteoblasts can promote chondrocytes migration, enhance gene expression of chondrocytes MMP-1, MMP-2, MMP-3, MMP-9 and regulate gene expression of TIMPs family. Interleukin-1β inhibitsthe migration of chondrocytes co-cultured with osteoblasts and gene expression of TIMPs family.

Objective To investigate the safety and therapeutic effects ot a novel multi-layer flat-plate bioartificial liver (BAL) for patients with liver failure.Methods Thirty-eight patients with liver failure from Dec.2010 to Dec.2011 were treated with a novel BAL based on multi-layer flatplate bioreactor and the co-cultured cells of the porcine hepatocytes and mesenchymal stem cells.A total of 48 treatments was performed,4 h each time.The clinical signs and symptoms,liver function,ammonia,coagulation function and complete blood count were evaluated before,during and after the treatment.DNA in the collected PBMCs was extracted for PCR with PERV specific primers and the porcine specific primer Sus scrofa cytochrome B.The RT activity was detected as well.Levels of xenoantibodies (IgG,IgM) were determined by using ELISA kit. Titers of complement were quantified by CH50 kit.Results All treatment procedures were completed successfully without any adverse reaction. All samples presented negative PERV DNA and RT activity. The levels of antibodies were similar before and after treatment.Treatment was associated with a temporary decline in levels of complement,and then the levels were recovered quickly.The clinical symptoms such as acratia,anorexia and abdominal distension were improved.The stage of hepatic encephalopathy in 16 patients was decreased. The liver function and ammonia was reduced disproportionately. Seven patients in all were bridged to liver transplantation,2 patients died and 2 patients gave up the treatment,and the others were turned better.After the outcome judgment according to the standard developed by the Artificial Liver Group,and Chinese Association of Infectious and Parasitic Diseases,there were 9 patients with clinical healing,25 patients with improvement and 4 patients with no effect,and the cure-improvement rate was 89.5％.Conclusion The novel multi-layer flat-plate BAL could be used as a safe and effective therapy for patients with liver failure.

Objective:To analyze the expression and role of activated transcription factor 3 (ATF3) in breast cancer.Methods:The Cancer Genome Atlas\Gene Expression Omnibus (TCGA\GEO) database information was queried, the expression level of ATF3 in breast cancer was analyzed by bioinformatics method and the survival curve was drawn. the parallel function and pathway enrichment analysis of ATF3 gene interaction network were constructed.Results:The level of ATF3 mRNA in breast cancer was significantly lower than that in normal breast tissue, but the expression level of ATF3 was significantly increased after chemotherapy. Survival analysis found that after systematic treatment, the survival period of breast cancer patients in the ATF3 high expression group was significantly prolonged, and the difference was statistically significant ( P<0.05). Enrichment analysis revealed that breast cancer-related pathways concentrate in Estrogens signaling pathway, mitogen-activated protein kinase kinases (MAPK) signaling pathway, phosphatidylinositol 3-kinase-protein kinase B (PI3K-Akt) signaling pathway, Wnt signaling pathway, Breast cancer signaling pathway, and Cell cycle signaling pathway. Conclusions:ATF3 is involved in regulating the proliferation of breast cancer cells. After systematic treatment, the high expression of ATF3 indicates a better outcome.

Objective To investigate the gene expression of Piezo1 in four types of bone cells at different stages of osteogenic differentiation under fluid shear stress (FSS). Methods The mouse-derived mesenchymal stem cells (MSC), osteoblast-like cells MC3T3-E1, post-osteoblasts MLO-A5 and osteocytes MLO-Y4 were exposed to FSS at different magnitude (0.1, 1.1 Pa) with a custom-made cone-plate flow chamber for 0.5, 1, 3, 6, 12 h, respectively. The expression of Piezo1 mRNA was assessed by quantitative real-time polymerase chain reaction. Results Both Piezo1 and Piezo2 were expressed in four types of bone cells. The expression of Piezo1 was significantly up-regulated in all cells under FSS stimulation, and the expression level under 1.1 Pa FSS was significantly higher than that under 0.1 Pa FSS. In addition, the expression of Piezo1 in MSC, MC3T3-E1 and MLO-A5 cells increased to the highest level at 1 h under FSS stimulation. The expression of Piezo1 in MC3T3-E1 cells was much higher than that in the other three types of cells. Conclusions The expression of Piezo1 was related to the process of osteogenic differentiation, FSS level and loading time, and this research finding is of great significance to reveal the mechanism of mechanotransduction in bone tissues and to establish clinical treatment for bone diseases.

Objective: To explore the topological abnormality of brain metabolic network in patients with idiopathic rapid eye movement sleep behavior disorder (iRBD) and compare it with the topology of brain metabolic network in patients with Parkinson′s disease (PD). Methods: The ¹⁸F-fluorodeoxyglucose (FDG) PET brain images of 19 patients with iRBD diagnosed with polysomnography (PSG) (iRBD group; 15 males, 4 females, average age: 64.9 years), 19 patients with PD (PD group; 12 males, 7 females, average age: 62.2 years) and 19 gender and age-matched healthy controls (HC group; 15 males, 4 females, average age: 63.1 years) in Huashan Hospital from September 2014 to June 2015 were retrospectively analyzed. According to the complex brain network method based on graph theory, the brain metabolic networks of each group was constructed and the network parameters (clustering coefficient, characteristic path length, local efficiency, global efficiency and small-world property, etc) were evaluated quantitatively. The 500 times non-parametric permutation test was used to determine the differences in network parameters between groups. Results: The brain metabolic networks of iRBD group and PD group both had abnormal topological structure, which showed that the characteristic path length (for example, when sparsity=34%, HC vs iRBD vs PD groups: 1.517 vs 1.552 vs 1.561) and local efficiency (for example, when sparsity=30%, HC vs iRBD vs PD groups: 0.802 vs 0.824 vs 0.831) were significantly increased (both P<0.05), the global efficiency (for example, when sparsity=36%, HC vs iRBD vs PD groups: 0.672 vs 0.658 vs 0.656) was significantly decreased (P<0.05). The topology was more aggravated in PD group compared with that in iRBD group. Conclusion The graph-based complex brain network analysis can reveal the abnormal topological structure of the brain metabolic network in which iRBD progresses to PD.