Nucleotide-binding oligomerization domain protein 2 (NOD2) involves in host immune responses to pathogens and regulates natural immunity and specific immunity by identifying the peptidoglycan of bacterial cell walls and cleavage product muramyl dipeptide.As a newly discovered intracellular pattern recognition receptor,NOD2 plays important roles in the development of primary hepatocellular carcinoma by gene mutate,inducing liver inflammatory reaction and activating relevant signaling pathways.

Objective To investigate galectin3 on proliferation and migration of esophageal cancer Eca109 cells.Methods A lentiviral vector for over-expression of RNA targeting galectin3 was designed to transfect Eca109 cancer cells following plasmid-mediated transfection manual (Eca109/Gal3 cells).Inverted fluorescence microscope was used to observe the expression of EGFP.The proliferation of Eca109 cells was measured by cell counting Kit-8 assay.Eca109 cells apoptosis was determined by Annexin-V/7-AAD doublestaining.The migration capacity of Eca109 cells was determined in transwell assays.Western blot analysis was used to measure the expression of galectin3 protein.Results Galectin-3 expression was detected in Eca109 cells,with the Galectin3 expression in Eca109/Gal3 cells much more than non-transfected cells (t =14.33,P ＜ 0.05 ; t =10.28,P =0.037).Compared with non-transfected Eca109 cells,proliferation increased significantly in Eca109/Gal3 cells (t =-17.277,P ＜ 0.05 ; t =-13.4,P ＜ 0.05).Galectin3 evidently decreased in Eca109 cell apoptosis (t =3.053,P ＜ 0.05 ; t =5.446,P ＜ 0.05).Transwell migration assay showed that a greater number of Eca109/Gal3 cells crossed the artificial basement membrane compared with non-transfected Eca109 cells and negative control Eca109 cells (t =3.465,P ＜ 0.05; t =3.252,P ＜ 0.05).Conclusion Galectin3 expression is detected in transfected esophageal cancer Eca109 cells,whose overexpression can result in enhanced proliferation,migration,invasion as well as reduced apoptosis.These data indicate that in-depth research of galectin-3 may prove to be a potential molecular target for the treatment of esophageal cancer.

Objective To explore the correlation of anti-ganglioside antibodies with clinical features of systemic lupus erythematosus (SLE). Methods Samples of serum were collected from 48 patients with SLE. Three were male and 45 were female. Their average age was 33±12 (15～59). The serum anti-ganglioside antibodies of 10 patients with rheumatoid arthritis (RA), 4 with Sj(o)gren's syndrome (SS), 1 patient with mixed connective tissue disease (MCTD), 1 with polymyositis (PM), and 97 healthy controls(HC) were alsoexamined. Levels of anti-GM1-IgM/IgG, anti-GQ1b-IgM/IgG antibodies in the serum were examined by amodified ELISA. Anti-dsDNA-IgG antibody of SLE was tested according to the ELISA kit protocol. The associations of anti-GM1-IgM/IgG and anti-GQ1b-IgM/IgG antibodies with clinical features were analyzed. x2test, one-way ANOVA, Dunnett t-test were used for statistical analysis. Results By using modified ELISA,the upper limit of reference value of anti-GM1-IgM/IgG antibodies, anti -GQ1b-IgM/IgG antibodies in the serum of Chinese SLE patients were 0.411, 0.408, 0.481 and 0.441 res-pectively. 19% patients with SLE were sero-positive for anti-GM1 antibody, 4% for IgM antibody isotype and 15% for IgG antibody isotype. Anti-GM1-IgG antibody was significantly increased in the serum of patients with SLE (0.33±0.09), higher than that of the HC (0.27±0.05) (P<0.05), RA (0.29±0.08), SS(0.27±0.06), PM ( 0.288 ), but one of the MCTD( 0.423 ).There was no significant associations between anti-GM1-IgM/IgG, anti-GQ1b-IgM/IgG antibodies and NPSLE and anti-dsDNA-IgG antibody (P>0.05). Conclusion Peripheral autoimmune response against GM1 can be detected in SLE patients, and it may be involved in the pathogenesis of SLE. No association of antiGM1-IgM/IgG antibodies or anti-GQ1b-IgM/IgG antibodies with clinical features of SLE can be discovered.

Objective To investigate the effect and mechanism of anti-tumor necrosis factor (TNF)-α on the intestinal mucosal permeability in dextran sulfate sodium (DSS) induced colitis mice.Methods Eighteen C57BL/6J mice were evenly divided into healthy control group,model group and anti-TNF-α treated group.The mice of model group and anti-TNF-α treated group were fed with 5％DSS solution for 7 days.The mice of anti-TNF-α treated group were injected anti-TNF-α (5 mg/kg)intraperitoneally on the first and fourth day; control group and model group were substituted with equal volume saline injection.The mice were sacrificed at 7 days after modeling.The disease activity index (DAI) score was evaluated everyday.The intestinal permeability was examined by Evan′s blue (EB) method and FITC-dextran (FITC-D) method.The colon tissue was collected for observation under microscope and histological index (HI).The small intestinal tissues were examined under electron microscope.The 10％ homogenate of colon and intestinal mucosa was prepared,the activity of myeloperoxidase (MPO),the content of TNF-α and epithelial myosin light chain kinase (MLCK) concentration were determined with kits respectively.The expression of MLCK in intestinal mucosa was tested by Western blot assay.Single factor of variance between groups were analyzed.Results Compared with control group,the DAI of model group increased daily.Compared with model group,the DAI of anti-TNF-α treated group improved.In model group,mice intestinal epithelial cells junctional complex shortened and widened and the cell gap expanded.In anti-TNF-α treated group,the connection structure of mice intestinal epithelial cells was tighter.The activity of HI and MPO and the content of TNF-α of model group were higher than those of control group (P ＝ 0.008,0.006 and 0.001,respectively),all of those of anti-TNF-α treated group were lower than those of model group (P=0.004,0.008 and 0.005,respectively).The F value of three groups was 131.98,218.28 and 58.93,respectively.The contents of EB in mice intestinal wall and serum FITC-D of model group were higher than those of control group (P=0.003 and 0.010),and those of anti-TNF-α treated group were lower model group (P＝0.001 and 0.009).The F value of three groups was 69.36 and 17.96.The MLCK concentration in mice intestinal mucosa of model group [(71.10± 7.52) ng/g] was higher than that of control group [(18.56±9.92) ng/g,P＜0.01],that of anti-TNF-α treated group [(37.56±15.84) ng/g] was lower than model group (P=0.008),and the difference among these three groups was statistically significant (F＝ 17.23).The Western blot results indicated the expression of MLCK in intestinal mucosa of model group was higher than that of control group,and that of anti-TNF-α treated group was lower than model group.Conclusions Anti-TNF-α play an important role in improving colitis,and the intestinal mucosal permeability.The mechanism may be related with the regulation of MLCK expression.

Animals ; Rats ; Specimen Handling ; methods ; Spinal Cord

Objective To investigate the expression of nucleotide-binding oligomerization domain protein 2 (NOD2)in serum of patients with hepatocellular carcinoma (HCC),and to analyse the roles of NOD2 in HCC development and its clinical diagnostic value.Methods This study including 66 patients with HCC in the hospi-tal from March 1,2013 to December 31,2014 and 61 healthy controls.Serum NOD2 levels were determined by enzyme-linked immunosorbent assay (ELISA).Analysis of significance was performed with rank sum test using SPSS statistical 16.0 software.Results Serum levels of NOD2 in HCC patients were 171 pg/ml,significantly higher than that of healthy controls(95 pg/ml,Z =-5.00,P =0.00),and the serum NOD2 levels were correla-ted with clinical stage of HCC (H=56.26,P =0.00).Compared with the serum NOD2 levels in stageⅠ,Ⅱpatients (106 pg/ml)and healthy controls (95 pg/ml),the serum NOD2 level in stage Ⅲ and Ⅳ (220 pg/ml) were significantly increased (χ2 =31.24,P =0.00;χ2 =47.23,P =0.00),but the expression of NOD2 in stageⅠandⅡwere nearly equal to that of the healthy controls (χ2 =0.36,P =0.83).The ROC analysis revealed that the best diagnostic cutoff-point of serum NOD2 levels for predicting the Ⅲ and Ⅳ stages of HCC was 148.78 pg/ml,meanwhile corresponding sensitivity was 89.1% and specificity was 77.0%.Additionally,corre-lation analysis demonstrated that there was no significant correlation between NOD2 and alpha-fetal protein (r =0.44,P =0.14).Survival curves obtained that the survival time of HCC patients with NOD2 serum concentrations≥ 200 pg/ml was significantly less than that <200 pg/ml (χ2 =15.32,P <0.05).Conclusion NOD2 is highly expressed in the serum of HCC patients,especially in advanced patients,which is possibly involved in the development of HCC and has the potential to become an effective marker used for HCC diagnosis.

Objective:To explore the isolation and culture of a better yield of purified human endometrial glandular epithelial cells and stromal cells. Methods:Digestion with high concentrated collagenase and DNase,filtratinn and adhesion purification were used to isolate,purify human endometrial glandular epithelial cells and stromal cells in 30samples. And the two kinds of cells were reproduced. Results:The endometrial cell culture was successfully established in 29 of 30 endometrium samples. From one gramme of endometrial tissue,the yields of glandular epithelial cells and stromal cells were (40～50)?10~6 and (40～50)?10~6 respectively. The purities of glandular epithelial cells and stromal cells were 96% and 98% respectively. Stromal cells could be reproduced into several passage. However,glandular epithelial cells could not be reproduced. Conclusion:A better yield of purified human endometrial glandular epithelial cells and stromal cells can be obtained by the modified culture procedure.

Animals ; Disease Models, Animal ; Fatty Liver ; Mice ; Rats

Catalyzed by a family of enzymes called glycosyltransferases (GTases), glycosylation reactions are essential for the bioactivities of macrolide antibiotics which have been widely applied. Additionally, glycosylation is also an important strategy of microbial to get macrolide antibiotic resistance. Studies on the structure, function and application areas of macrolide GTases will lay the stable groundwork for the combinatorial biology. This paper introduced in detail the biological functions of macrolide glycosylation, and then made an in-depth discussion on the families and discoveries of macrolide GTases. The resistance mechanism with macrolide glycosyltion and the correlative GTases MGT have been reviewed afterwards. According to the flexible substrate specificity of macrolide GTases, the combinatorial biological applications on them were also seriously summarized here. At the end, the authors made a developmental prospect of macrolide GTases based on the studies of the research group.
Anti-Bacterial Agents ; metabolism ; pharmacology ; Drug Resistance, Bacterial ; Glucosyltransferases ; classification ; metabolism ; Glycosylation ; Macrolides ; chemistry ; metabolism ; Streptomyces ; enzymology ; Substrate Specificity

In tumor immunotherapy, the study of programmed death 1 (PD-1) and programmed death ligand 1 (PD-L1) provides a new approach for the comprehensive treatment of advanced lung cancer. About 20％ of patients with non-small cell lung cancer could benefit from lung cancer immunotherapy, while those with high PD-L1 expression will benefit more. At present, there are few related studies on PD-1 expression at home and abroad, and the detection of PD-1/PD-L1 expressions is mostly concentrated in tumor tissues. With the research progress of liquid biopsy technology, the convenience and accuracy of peripheral blood testing are also receiving more and more attention. However, there are still few biological indicators for predicting the efficacy of tumor immunotherapy, and the uniform standard and accuracy of testing still need more clinical practice and exploration. This article reviews the research on the expressions of PD-1 and PD-L1 in tissues and peripheral blood of patients with lung cancer, aiming to provide reference for the treatment of lung cancer with immune checkpoint blockers.