Expression of Far Upstream Element Binding Protein 1 in Gastric Cancer and Its Effect on Proliferation and Invasion of Gastric Cancer Cells / 中国医科大学学报

Objective To investigate the expression of far upstream element binding protein 1 (FUBP1) gene in gastric cancer,and analyze its effect on proliferation and invasion of gastricc cancer cells.Methods The expression of FUBP1 in gastric cancer tissues was detected by real-time PCR.The relationship between FUBP1 expression and clinic pathological factors of gastricc cancer was analyzed by one-way ANOVA.The silence vector for FUBP1 was constructed and transfected into gastricc cancer SGC7901 cells,and the transfected effects were then detected by real-time PCR.The influence of FUBP1 silence on cell proliferation and invasion in SGC7901 cells were detected by CCK8 method and Transwell assay.Results FUBP1 expression was increased markedly in gastricc cancer tissues.Following the depressing of differentiation and the increase of invasion depth and metastatic lymph nodes,the FUBP1 expression was up-regulated in primary gastric carcinoma significantly.The pS-FUBP1 vector could silence the FUBP1 expression in SGC7901 cells.FUBP1 silence inhibited SGC7901 cell proliferation amd invasion.Conclusion FUBP1 was high-expressed in gastricc cancer,and related with genesis and progress of gastric cancer,silence of FUBP1 expression can inhibit the cell proliferation and invasion in gastric cancer cells.

Natural killer cell promotes the HBV clearance via secreting IFN-γ / 安徽医科大学学报

Objective To investigate the mechanism of NK cell in eliminating the Hepatitis B virus during HBV in-fection . Methods Acute HBV infection model was established by injecting adult mice hydrodynamically with 20μg of pGEM4Z/HBV1. 2 plasmid. This model was evaluated by detecting serum level of HBsAg and HBcAg in liver tis-sue at the indicated time points by radioimmunoassay and immunohistochemistry respectively. The variation of fre-quency and absolute number of NK cell was analyzed between wide type ( WT ) mice and HBV plasmid-injected mice. Furthermore, the activation and the IFN-γproduction of NK cell were investigated in these mice by flow cy-tometry. HE staining and alanine transaminase( ALT) dectection were used to observe liver injury. To test whether NK cell and IFN-γwere involved in HBV elimination, we used PK136 antibody to clear NK cell and IFN-γneutral-ization antibody toblock IFN-γeffect. Results After the hydrodynamic injection with 20 μg of pGEM4Z/HBV1.2, the serum level of HBsAg and expression of HBcAg in liver tissue were very high at 1 week, but then decreased gradually. However, these antigens almost became negative at 4 to 5 weeks, which mimic acute HBV infection pa-tients. Compared with NK cell from WT mice, the frequency and absolute number of NK cell increased significantly from HBV mice. Also, the NK cells express higher level of CD69 and produce more IFN-γ. Meanwhile, there was no liver injury in HBV mice. Depletion of NK cell or blocking IFN-γ effect in HBV mice could significantly in-crease the level of HBV related antigens. Conclusion In the mouse model of acute HBV infection, NK cell could promote the HBV elimination through secreting IFN-γ.

Clinical analysis of common abdominal operation complicated with pleural effusion / 国际外科学杂志

Pleural effusion is one of the common complications after abdominal surgery, which has a high incidence rate. It not only extends the length of hospital stay and increases the economic burden, but also may affect the prognosis of patients or even endanger lives. Therefore, early diagnosis and treatment of pleural effusion are important for postoperative rehabilitation. So far,the relationship between pleural effusion and clinical pathological factors of postoperation has not been revealed clearly and definitely. Preventive measures and the causes are summarized by reading a large number of literatures and combining with clinical experience to reduce incidence rate.

Lipopolysaccharide induces parkin expression and mitophagy in murine peritoneal macrophages / 中国免疫学杂志

Objective: To investigate whether lipopolysaccharide induced parkin expression and mitophagy in macrophages.Methods:The murine peritoneal primary macrophages were aseptically isolated from Kunming mice and cultured in complete medium.The mitochondrial membrane potential of macrophages was detected by flow cytometry,after the cells were stimulated with 200 ng/ml LPS and labeled mitochondria with JC-1.The parkin mRNA level of macrophages was detected by RT-PCR, protein levels of parkin and autophagic related protein LC3 Ⅱ and LC3 Ⅰ were determined by Western blot.The distribution and co-localization of parkin with LC3 and mitochondria in macrophages were respectively observed by laser scanning confocal microscope, before and after the cells were treated with LPS.Results: Flow cytometry results after JC-1 staining showed that mitochondrial membrane potential in macrophages was declined after stimulation with 200 ng/ml LPS, and continuously decreased with prolonged treatment time.The mRNA levels of parkin were increased slightly within 6 h after LPS stimulation,but parkin proteins were increased significantly within 6 h after LPS stimulation.The results of parkin distribution showed that parkin was evenly distributed in the cytoplasm at normal status, but became the obvious punctate distribution after LPS stimulation in macrophages.Western blot results showed LC3 Ⅱ/LC3 Ⅰ levels were increased after LPS stimulation, indicating the appearance of macrophage autophagy.Confocal microscopy showed that there were co-localization of parkin,LC3 and mitochondrial in macrophages after LPS stimulation.Conclusion:Parkin expression is increased significantly and mediated mitochondrial autophagy in macrophages after LPS stimulation, which is involved in the clearance of damaged mitochondria,thereby playing a role in regulating macrophage inflammatory response.

Effect of simvastatin on production of reactive oxygen species and secretion of IL-1βin macrophages induced by oxLDL / 中国药理学通报

Aim To study the effect of simvastatin on the production of reactive oxygen species ( ROS ) and the secretion of interleukin-1 beta ( IL-1β) in oxidized low density lipoprotein ( oxLDL )-induced macropha-ges. Methods After the murine macrophage J774A. 1 was treated with 0,50,100,200 mg·L-1 oxLDL, the contents of aggregated lipid in macrophages were ob-served and determined by oil red O staining. Then, the oxLDL-primed macrophages were treated with 0 . 5 ,1 . 0μmol·L-1 simvastatin, the production of ROS was de-termined by flow cytometry and the expressions of pro-caspase-1 , cleaved caspase-1 and mature IL-1βon pro-tein level were determined by Western blot. Results The oil red O staining results showed that oxLDL could induce obvious lipid aggregation in macrophages, and reached the saturation point with 100 mg·L-1 concen-tration. Flow cytometry results indicated that oxLDL could induce the production of ROS in macrophages, up to 167% ± 0. 47%, and ROS level decreased to 139% ± 0. 97% in a dose-dependent manner after treatment with simvastatin. Western blot indicated that simvastatin could inhibit the expression of cleaved caspase-1 and mature IL-1β in macrophages triggered by oxLDL;compared with oxLDL group, the expression of cleaved caspase-1 and mature IL-1β decreased in simvastatin treated group, and all results had statistical significance ( P<0. 05 ) . Conclusion In the lipid ag-gregation model of macrophages induced by oxLDL, simvastatin can inhibit the production of ROS, caspase-1 activation, and secretion of IL-1β in macrophages.

Preparation and Characterization of Butylaminopropyl Silica Monolithic Column for Capillary Electrochromatography / 分析化学

A novel silica monolithic stationary phase functionalized with butylaminopropyl ligands for capillary electrochromatography(CEC) has been presented. The monolithic capillary columns were prepared by a sol-gel) process and subsequent a chemical modification. The amino groups on the surface of the stationary phase are meant to generate a substantial anodic electroosmotic flow (EOF). The butyl and propyl groups provide) hydrophobic properties. To evaluate the column performance, effects of buffer pH and organic modifier content on the EOF and electrochromatographic retention behavior of alkylbenzenes, organic acids and anilines were investigated. The monolithic stationary phase exhibited reversed phase (RP) chromatographic behavior toward neutral solutes. The model organic acid anion solutes were separated by the mixed mode mechanism, which comprised RP interaction, weak anion-exchange, and electrophoresis. Basic compounds such as anilines) were well separated on the butylaminopropyl silica monolithic column without peak tailing.

Donor heart gene transfer of CD40Ig mediated by adenovirus to prolong the survival time of murine cardiac allograft / 中华器官移植杂志

Objective To explore the effects of CD40Ig gene transfer in vitro on cardiac allograft survival in mice. Methods The recombinant adenovirus vector carrying murine CD40 extracellular domain and human IgG Fc fusion gene (AdCD40Ig) was constructed. Using BALB/c mice as donors and C57BL/6 mice as recipients, the model of mice abdomen heterotopical heart transplantation was set up. In the experimental group, the isolated donor heart was transfected with AdCD40Ig, then transplanted to recipient. The other groups included empty vector transfected control group, untransfected control group, and syngeneic control group (the donors and the recipients were C57BL/6 mice). The survival time of donor hearts and the infiltration of inflammatory cells in heart allograft were observed. The expression of CD40Ig fusion protein in recipient was identified by Sandwich ELISA; The IFN-? producing cells in recipient was determined by FACS. Results The survival time of the cardiac allograft in experimental group was ( 15.8? 0.7) days, significantly longer than empty vector transfected control group and untransfected control group (P

Aberration of CCR7 CD8 memory T cells from patients with systemic lupus erythematosus: an inducer of T helper type 2 bias of CD4 T cells.

Chemokine receptors are important in the entry of leucocytes into the inflammatory sites of systemic lupus erythematosus (SLE). CCR7(+) and CCR7(-) memory T cells exert different functions in homing, cytokine production and cytotoxicity. To determine whether differential expression and functions of the CCR7 occur in SLE patients, we examined CCR3, CCR4, CCR5, CCR7 and CCR9 on CD4(+) and CD8(+) T cells from normal and SLE subjects. Flow cytometry, real-time quantitative reverse transcription polymerase chain reactions and Northern blotting were used to detect the expression of chemokine receptors and cytokines; a chemotaxis assay was used to detect their functions. CD4(+) T-cell stimulation with syngeneic CCR7(+) CD8(+) CD45RO(+) T cells and dendritic cells (including transwell chambers) was used to induce cytokine expression. We demonstrated that CCR7 was selectively, frequently and functionally expressed on CD8(+) (94.8%) but not on CD4(+) (16.1%) T cells from patients with active SLE, whereas this phenomenon was not seen in normal subjects and in those whose SLE was inactive. CCR7(+) CD8(+) CD45RO(+) memory T cells from patients with active SLE, themselves T helper type 2 (Th2) biased, were inducers of Th2 bias in CD4(+) T cells in a cell-cell contact manner in vitro, meanwhile, the cells from both normal subjects and those whose SLE was inactive drove CD4(+) T cells into a regulatory T-cell-derived cytokine pattern. Our findings might provide new clues to understanding the functions of CCR7(+) CD8(+) CD45RO(+)'central' memory T cells in autoimmune diseases (such as SLE). We suggest that in the case of active SLE, CCR7(+) central memory T cells were able to enter peripheral blood and inflammatory sites from secondary lymphoid organs, were continuously expressing CCR7, and interacted with dendritic cells and functioned as CCR7(-)'effector' memory T cells, which were described in normal humans.

Highly up-regulated CXCR3 expression on eosinophils in mice infected with Schistosoma japonicum.

CXCR3, predominately expressed on memory/activated T cells, is a receptor for both interferon-gamma inducible protein-10/CXC ligand 10 (CXCL10) and monokine induced by interferon-gamma/CXCL9. We reported here that CXCR3 was highly up-regulated on infiltrating eosinophils in Schistosoma japonicum egg-induced granuloma in the mouse liver. It was also highly and functionally up-regulated on peritoneal exudate eosinophils in mice infected with S. japonicum. The phenomena were demonstrated at protein and mRNA levels using immunohisto- and immunocytochemistry evaluation of biopsy, flow cytometry and real-time quantitative reverse transcriptase-polymerase chain reaction technique, and verified by Northern blotting and chemotaxis assay in vitro. We also found that CCR3 expression on the infiltrating and peritoneal exudate cells was significantly decreased, CXCR4 expression was unchanged during the 42-day period of infection. We screened mRNA expression levels of the all known chemokine receptors in purified peritoneal exudate eosinophils and liver granuloma dominated by eosinophils. CXCR3 was highly and functionally up-regulated on peritoneal exudate eosinophils in mice infected with S. japonicum, meanwhile CCR3 was significantly and functionally down-regulated in these cells. The findings could lead to a better understanding of the chemokine receptor expression pattern of eosinophils at inflamed tissue sites caused by parasites. These could be also crucial for establishing a therapeutic strategy for eosinophilic inflammation via intervention in chemokine actions.

Selectively increased expression and functions of chemokine receptor CCR9 on CD4+ T cells from patients with T-cell lineage acute lymphocytic leukemia.

In a total of 38 typical T-cell lineage acute lymphocytic leukemia (T-ALL) and T-cell lineage chronic lymphocytic leukemia (T-CLL) cases investigated, we found that CC chemokine receptor CCR9 was selectively and frequently expressed on T-ALL CD4+ T cells, was moderately expressed on T-CLL CD4+ T cells, and was rarely expressed on normal CD4+ T cells. These findings were demonstrated at protein and mRNA levels using flow cytometry and real-time quantitative reverse transcription-PCR technique and were verified by digital confocal microscopy and Northern blotting. Thymus-expressed chemokine, a ligand for CCR9, selectively induced T-ALL CD4+ T-cell chemotaxis and adhesion. Interleukin (IL)-2 and IL-4, together, down-regulated the expression and functions of CCR9 in T-ALL CD4+ T cells including chemotaxis and adhesion. It was also demonstrated that IL-2 and IL-4, together, internalized CCR9 on T-ALL CD4+ T cells and subsequently inhibited functions of CCR9 in these cells. Thymus-expressed chemokine mRNA was highly expressed in CD4+ T cells, involving lymph node and skin in T-ALL patients, and was expressed at moderate levels in lymph node and skin tissues in T-CLL patients. Our findings may provide new clues to understanding various aspects of T-ALL CD4+ T cells, such as functional expression of CCR9-thymus-expressed chemokine receptor-ligand pairs as well as the effects of IL-2 and IL-4, which may be especially important in cytokine/chemokine environment for the pathophysiological events of T-ALL CD4+ T-cell trafficking.

Peptide nucleic acid antisense prolongs skin allograft survival by means of blockade of CXCR3 expression directing T cells into graft.

CXCR3, predominantly expressed on memory/activated T cells, is a receptor for both IFN-gamma-inducible protein 10/CXC chemokine ligand (CXCL)10 and monokine induced by IFN-gamma/CXCL9. It was reported that CXC chemokines IFN-gamma-inducible protein 10/CXCL10 and monokine induced by IFN-gamma/CXCL9 play a critical role in the allograft rejection. We report that CXCR3 is a dominant factor directing T cells into mouse skin allograft, and that peptide nucleic acid (PNA) CXCR3 antisense significantly prolongs skin allograft survival by means of blockade of CXCR3 expression directing T cells into allografts in mice. We found that CXCR3 is highly up-regulated in spleen T cells and allografts from BALB/c recipients by day 7 of receiving transplantation, whereas CCR5 expression is moderately increased. We designed PNA CCR5 and PNA CXCR3 antisenses, and i.v. treated mice that received skin allograft transplantations. The PNA CXCR3 at a dosage of 10 mg/kg/day significantly prolonged mouse skin allograft survival (17.1 +/- 2.4 days) compared with physiological saline treatment (7.5 +/- 0.7 days), whereas PNA CCR5 (10 mg/kg/day) marginally prolonged skin allograft survival (10.7 +/- 1.1 days). The mechanism of prolongation of skin allograft survival is that PNA CXCR3 directly blocks the CXCR3 expression in T cells, which is responsible for directing T cells into skin allograft to induce acute rejection, without interfering with other functions of the T cells. These results were obtained at mRNA and protein levels by flow cytometry and real-time quantitative RT-PCR technique, and confirmed by chemotaxis, Northern and Western blot assays, and histological evaluation of skin grafts. The present study indicates the therapeutic potential of PNA CXCR3 to prevent acute transplantation rejection.

The expression and clinical significance of multi-genes in breast cancer / 中国普通外科杂志

0.05). Expression of MDR1 had a positive ~correlation with mutant p53 accumulation and HER2 expression(P0.05 ).In univariate analyses,TNM staging, axillary lymph node metastasis, mutant p53 accumulation, and HER2 over-expression were negatively correlated with DFS and OS, and MDR1 over-expression significantly reduced OS but not DFS. In multivariate analysis, axillary lymph node metastasis, over-expression of MDR1 and HER2 were independent risk factors for prognosis. Conclusions ~Induction of multidrug resistance and poor response to chemotherapy and endocrinotherapy may be the chief reasons for poor prognosis of breast cancer with mutant p53 accumulation, and HER2 and MDR1 over-expression. ~Determination of the above genes′expression in breast cancer tissue can be of use in deciding the degree of ~malignancy , metastasis phenotype and prognosis of brest cancer. Increasing anthracycline dose may increase the ~overall response rate to chemotherapy and improve prognosis in patients with mutant p53 accumulation, HER2 and MDR1 over-expression, especially HER2 over-expression.