OBJECTIVE: To investigate the significance of Tim-3 and Galectin-9 in Th1/Th2 imbalance in patients with multiple myeloma (MM). METHODS: 55 newly diagnosed MM patients and 20 healthy controls were included. Flow cytometry was used to detect the expression of Tim-3 on CD4⁺T cells, the proportion of Th1, Th2, Tim-3⁺Th1 and Tim-3⁺Th2 cells in peripheral blood. ELISA was used to detect the levels of cytokines IFN-γ and IL-4 in serum, and PCR was used to detect the level of Galectin-9 mRNA. Then the correlations between Galectin-9 mRNA expression and Th-cell subsets and related cytokine levels, as well as the relationship between Tim-3⁺Th1/Tim-3⁺Th2 ratio and corresponding clinical features were analyzed. RESULTS: Compared with the control group, the expression of Tim-3 on CD4⁺T cells in peripheral blood of MM patients was significantly increased (P<0.05), the proportions of Tim-3⁺Th1 cells, Tim-3⁺Th2 cells and Tim-3⁺Th1/Tim-3⁺Th2 ratio in MM patients were also increased (P<0.05), while the proportion of Th1 cells and Th1/Th2 ratio in MM patients were significantly decreased (P<0.05). The level of cytokine IFN-γ and IFN-γ/IL-4 ratio in MM patients were significantly decreased (P<0.05), while the level of cytokine IL-4 was increased (P<0.05). The mRNA levels of Galectin-9 in MM patients were significantly increased (P<0.05). The levels of Galectin-9 mRNA were positively correlated with Tim-3⁺CD4⁺T cells (r=0.663), Tim-3⁺Th2 cells (r=0.492) and IL-4 (r=0.470), while negatively correlated with IFN-γ (r=-0.593). The ratios of Tim-3⁺Th1/Tim-3⁺Th2 in MM patients were positively correlated with ISS stage (r=0.511), osteolytic damage (r=0.556) and chromosome abnormality (r=0.632). CONCLUSION These results suggest that Tim-3 and Galectin-9 are involved in Th1/Th2 imbalance in MM patients, and the high ratio of Tim-3⁺Th1/Tim-3⁺Th2 is associated with poor clinical prognosis.
Humans ; Cytokines/metabolism* ; Galectins/metabolism* ; Hepatitis A Virus Cellular Receptor 2/metabolism* ; Interleukin-4/metabolism* ; Ligands ; Multiple Myeloma/metabolism* ; RNA, Messenger/metabolism* ; Th1 Cells/metabolism* ; Th2 Cells/metabolism*

In order to prevent phage contamination in amino acid fermentation, we introduced the restriction and modification system cglI gene complex into Corynebacterium crenatum and studied their phage-resistance. The cglI gene complex was amplified from Corynebacterium glutamicum by PCR and constructed into pJL23 vector. The recombinant strains were obtained by transformation of the recombinant plasmid pJL23-cglI into C. crenatum. Results showed that the recombinant strains possessed strong phage-resistance activity and broad phage-resistance spectrum, demonstrating the feasibility of using cglI gene complex for construction of phage-resistance recombinant C. crenatum strains and presenting a powerful way to solve the problem of phage contamination in amino acid fermentation industry.
Amino Acids ; biosynthesis ; Bacterial Proteins ; genetics ; Bacteriophages ; growth & development ; Corynebacterium ; genetics ; virology ; Corynebacterium glutamicum ; genetics ; metabolism ; DNA Restriction-Modification Enzymes ; genetics ; Fermentation ; Galectins ; genetics ; Recombination, Genetic ; Transformation, Bacterial

OBJECTIVETo study the regulatory effect of galectin-9 isoforms on some molecules involved in cell adhesion/invasion, and the influence of this regulation action on the adhesion between colon carcinoma LoVo cells and human umbilical vein endothelial cells (HUVECs) in vitro.

METHODSVarious expression vectors of galectin-9 isoforms were transfected into LoVo cells. 24 h after transfection, the expression of integrin-beta1, E-cadherin, E-selectin, ICAM-1, CD44 and MMP-9 was detected by RT-PCR and Western blot analysis. LoVo cell-HUVEC adhesion assay was performed under conditions of galectin-9 transfection, galectin-9 transfection + galectin-9 antibody, galectin-9 transfection + E-selectin antibody and galectin-9 transfection + beta-lactose, respectively.

RESULTSGalectin-9L down-regulates the mRNA and protein levels of E-selectin while galectin-9M and galectin-9S up-regulate the expression of E-selectin. In LoVo cell-HUVEC adhesion assay, the average fluorescence intensity of vector transfection group, galectin-9L transfection group, galectin-9M transfection group and galectin-9S transfection group was 0.90 +/- 0.20, 0.94 +/- 0.24, 1.60 +/- 0.11 and 1.45 +/- 0.13, respectively, indicating that galectin-9M and galectin-9S facilitated the adherence of LoVo cells to HUVECs (P < 0.05). E-selectin antibody, galectin-9 antibody or beta-lactose inhibited that effect.

CONCLUSIONGalectin-9 isoforms regulate the E-selectin expression in LoVo cells differently and thus influence the adhesion between LoVo cells and HUVECs in vitro in different modes. The mechanisms through which galectin-9 isoforms participate in the metastasis process of colon cancer may not be the same.

Cell Adhesion ; Cell Line, Tumor ; Cells, Cultured ; Colonic Neoplasms ; metabolism ; pathology ; E-Selectin ; genetics ; metabolism ; Endothelial Cells ; cytology ; Galectins ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Humans ; Protein Isoforms ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Umbilical Veins ; cytology

OBJECTIVE: To observe the correlation between the expression of galectin-7 and S100A9 with the development of cervical squamous carcinoma. METHODS: Immunohistochemical SP staining was used to detect the expression of galectin-7 and S100A9 in 243 patients with cervical intraepithelial neoplasia (CIN) or cervical squamous carcinoma. The association of clinical data with galectin-7 and S100A9 expression was examined. RESULTS: The expression of galectin-7 and S100A9 in CIN and cervical squamous carcinoma was significantly different (P<0.05). The positive rates of galectin-7 in normal cervical tissues, CIN I, CIN II, CIN III, and cervical squamous carcinoma were 56.7%, 41.9%, 32.0%, 27.3%, and 25.0%, respectively. Statistic analysis found significant difference between the normal cervical tissues and cervical squamous carcinoma (P<0.0045). The positive rates of S100A9 in CIN I, CIN II, CIN III, and cervical squamous carcinoma were 80.0%, 77.4%, 48.0%, 27.3%, and 20.2%. Statistic analysis showed significant difference between the normal tissues and CIN III, the normal cervical tissues and cervical squamous carcinoma, CIN I and CIN III, CIN I and cervical squamous carcinoma, CIN II and cervical squamous carcinoma (P<0.0045). A positive correlation was found between galectin-7 and S100A9 expression in CIN and cervical squamous carcinoma (rs=0.298, P<0.001). Expressions of both galectin-7 and S100A9 in cervical squamous carcinoma were associated with the clinical stage and lymph nodes (P<0.05), but not with patient's age and degree of differentiation (P>0.05). Expression of galectin-7 was associated with the survival rate of patients with cervical squamous carcinoma (P<0.05). Univariate analysis of Cox proportional hazards regression model revealed that the FIGO stage, lymph nodes metastasis, and the expression of galectin-7 were relevant to the 5 year survival rate of patients with cervical squamous carcinoma, which was confirmed by multiple analysis of Cox proportional hazards regression model. CONCLUSION Expression of galectin-7 and S100A9 is related with cervical the tumorigenesis of carcinoma, clinical stage, and lymph nodes of cervical squamous carcinoma. Galectin-7 is probably associated with the prognosis. The long-term survival of patients with cervical carcinoma may be associated with FIGO stage, lymph node metastasis, and the expression of galectin-7.
Calgranulin B ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; Cell Differentiation ; Cervical Intraepithelial Neoplasia ; metabolism ; Female ; Galectins ; metabolism ; Humans ; Lymphatic Metastasis ; Prognosis ; Proportional Hazards Models ; Survival Rate ; Uterine Cervical Neoplasms ; metabolism

OBJECTIVETo construct the recombinant plasmid for Angiostrongylus cantonensis (AC) galectin (GAL) cDNA and analyze the immunological activity of the recombinant protein.

METHODSAcGAL cDNA was screened from the cDNA library and amplified by PCR. The amplified fragment was subcloned into the expression vector pET32a(+) and expressed in E.coli. The inclusion body was washed, degenerated, refolded by dialysis, and condensed for SDS-PAGE and Western blot analysis of the protein.

RESULTSFor the first time the full-length cDNA of AcGAL was cloned (GenBank GeneID: DQ384534). Restriction enzyme digestion indicated that the recombinant plasmid pET32a(+)-AcGAL was successfully constructed. SDS-PAGE analysis confirmed high expression of the recombinant protein AcGAL in E.coil in the form of inclusion bodies, which possessed good immunoreactivity as shown by Western blot analysis.

CONCLUSIONThe success in cloning and identification, the recombinant AcGAL may provide basis for further diagnostic study of angiostrongyliasis.

Angiostrongylus cantonensis ; genetics ; metabolism ; Animals ; Blotting, Western ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Galectins ; genetics ; metabolism ; Helminth Proteins ; genetics ; metabolism ; Molecular Sequence Data ; Recombinant Proteins ; metabolism ; Sequence Analysis, DNA

OBJECTIVETo evaluate the expression of galectin-3 and galectin-9 in mice nasal mucosa,and to explore the role of galectin-3 and galectin-9 in allergic rhinitis (AR).

METHODSTwenty mice were randomly divided into AR group and control group, 10 mice in each group. Ten mice of BALB/c were sensitized intraperitoneally with 10 microg of ovalbumin(OVA) adsorbed onto 2 mg Al(OH)3 on day 1 and day 14. Mice was induced daily by intranasal daily administration of 10 microl of saline containing 100 microg of OVA from day 21 to day 28. OVA was replaced with saline in control group. Immunohistochemical staining was used to examine the expression of galectin-3 and galectin-9 in nasal mucosa. RT-PCR was performed to investigate the level of mRNA expression of complement galectin-3, galectin-9. The level of IL-4, IL-5 and IFN-gamma in peripheral blood were titrated by ELISA.

RESULTSGalectin-3 and galectin-9 were detected in both groups. Expression of galectin-3 and galectin-9 in group AR was higher than that in control group. Pearson correlation analysis demonstrated that the levels of galectin-3 and galectin-9 were positively correlated with the level of IL-4 and IL-5, but negatively correlated with the level of IFN-gamma.

CONCLUSIONSGalectin-3 and galectin-9 might play an important role in the pathogenesis of allergic rhinitis.

Animals ; Female ; Galectin 3 ; metabolism ; Galectins ; metabolism ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Interleukin-5 ; blood ; Male ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; metabolism ; Rhinitis, Allergic, Seasonal ; metabolism

The relationship between T cell immunoglobulin domain and mucin domain protein 3 (Tim-3)/Galectin (Gal)-9 pathway and recurrent spontaneous abortion (RSA) was studied. Thirty-one pregnant women with RSA and 27 normal early gravidas were investigated to detect the levels of Tim-3 and Gal-9 in villi and deciduas by Western blotting. Meanwhile, the concentration of interleukin (IL)-4 and IL-12 in peripheral blood plasma was determined by ELISA in 25 healthy fertile non-pregnant controls, the normal early gravidas and pregnant women with RSA mentioned above, respectively. It was found that the relative expression levels of Tim-3 and Gal-9 in villi and deciduas were significantly increased in pregnant women with RSA as compared with those in the normal early gravidas. The concentration of IL-4 in peripheral blood plasma of pregnant women with RSA was lower than that of the normal early gravidas (P<0.05) and healthy fertile non-pregnant controls (P<0.05), but that of IL-2 in pregnant women with RSA was significantly higher than that of the normal early gravidas (P<0.05) and healthy fertile non-pregnant controls (P<0.05). It was suggested that the overexpression of Tim-3/Gal-9 pathway may be related to the pathogenesis of RSA.
Abortion, Spontaneous ; metabolism ; pathology ; Adolescent ; Adult ; Chorionic Villi ; metabolism ; pathology ; Female ; Galectins ; biosynthesis ; Hepatitis A Virus Cellular Receptor 2 ; Humans ; Interleukin-12 ; blood ; Interleukin-4 ; blood ; Membrane Proteins ; biosynthesis ; Pregnancy ; Pregnancy Proteins ; biosynthesis ; Up-Regulation

OBJECTIVEIt is supposed that bronchial epithelial cells responses to the environmental stimuli are different between asthmatic and non-asthmatic individuals, which contribute to the pathogenesis of asthma. These different responses produce different mediators. If differential gene expressions are found in bronchial epithelial cells of asthmatic and non-asthmatic individuals after the same stimuli in vitro, and these genes are overexpressed in asthmatic children in vivo, then it is concluded that these genes may be associated with asthma. Therefore the authors analyzed the differential gene expressions in the bronchial epithelium cells of asthmatic and non-asthmatic children after RSV infection in vitro. Among these genes, Galectine-7 (lectin, galactoside-binding, soluble, 7, Galectin-7) was 8 times up-regulated in asthmatic children. Galectine-7 was associated with skin keratinocyte apoptosis. The authors hypothesized that Galectin-7 may also be associated with bronchial epithelial cell apoptosis in asthmatic children. The aim of this study was to understand the role of Galectine-7 in bronchial epithelial cell apoptosis in asthma.

METHODSThe bronchial mucosae of one asthmatic child and one non-asthmatic child were obtained by biopsy and cultured in vitro. The bronchial epithelial cells were infected by RSV. The differential gene expressions were analyzed with micro array. Among those differentially expressed genes, Galectin-7 was 8 times up-regulated in asthmatic children. The bronchial mucosae from 10 asthmatic children and 17 non-asthma children were investigated for cell DNA break, Galectine-7 and mRNA expression, Caspase-3 expression by TUNEL, hybridization in situ and immunochemistry. Image analysis was used for quantitative assessment.

RESULTSGalectine-7 gene was 8 times up-regulated in bronchial epithelial cells from asthmatic children after RSV infection in vitro. Galectin-7 and mRNA were overexpressed in bronchial epithelial cells in asthma in vivo. Bronchial epithelial cell apoptosis increased in asthma in vivo.

CONCLUSIONGalectin-7 may be associated with bronchial epithelial cell apoptosis in asthma.

Adolescent ; Apoptosis ; genetics ; Asthma ; metabolism ; pathology ; Biopsy ; Bronchi ; metabolism ; pathology ; Bronchoscopy ; Caspase 3 ; genetics ; metabolism ; Cells, Cultured ; Child ; Child, Preschool ; Epithelial Cells ; metabolism ; pathology ; virology ; Female ; Galectins ; genetics ; metabolism ; Gene Expression Profiling ; Humans ; Immunohistochemistry ; In Situ Hybridization ; In Situ Nick-End Labeling ; Infant ; Male ; RNA, Messenger ; Respiratory Mucosa ; cytology ; metabolism ; pathology ; virology ; Respiratory Syncytial Viruses ; pathogenicity ; Up-Regulation

OBJECTIVETo study the expression of Galectin-9 and Tim-3 in lungs of mice with asthma and the effect of rosiglitazone (PPAR-γ agonist) on their expression.

METHODSFortyfive BALB/c SPF female mice were randomized into control group and asthma groups with and without rosiglitazone intervention. After ovalbumin stimulation and rosiglitazone intervention the pathological changes of the lung tissues were observed. Galectin-9 and Tim-3 mRNA levels in lung tissues were determined using RT-PCR. The levels of IL-4 and IFN-γ in peripheral blood were measured using ELISA.

RESULTSThe expression of Galectin-9 and Tim-3 mRNA of lung tissues in the untreated asthma group increased significantly compared with the control and the rosiglitazone treated groups (P<0.05). A significantly increased blood expression of IL-4 and a significantly decreased blood expression of IFN-γ were found in the untreated asthma group compared with the control and the rosiglitazone-treated groups (P<0.05). The expression of Galectin-9 and Tim-3 mRNA was positively correlated with blood IL-4 level (r=0.792, r=0.794 respectively; P<0.05), but negatively correlated with blood IFN-γ level (r=-0.692, r=-0.757 respectively; P<0.05).

CONCLUSIONSGalectin-9 and Tim-3 mRNA levels in lungs increase in mice with asthma and significantly correlate with the levels of blood Th1/Th2 cytokines. This suggests that Galectin-9 and Tim-3 are closely related to inflammatory process in asthma. Rosiglitazone treatment may decrease the expression of Galectin-9 and Tim-3.

Animals ; Asthma ; drug therapy ; immunology ; pathology ; Female ; Galectins ; genetics ; Hepatitis A Virus Cellular Receptor 2 ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Lung ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; PPAR gamma ; physiology ; RNA, Messenger ; analysis ; Receptors, Virus ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Thiazolidinediones ; therapeutic use