Emticicia soli sp. nov., a novel member of the family 'Flexibacteraceae', isolated from tetrabromobisphenol A-contaminated soil.

Bacterial strain ZZ-4T, a Gram-stain-negative, aerobic, non-spore-forming, non-motile, non-flagellated, rod-shaped bacterium, was isolated from tetrabromobisphenol A-contaminated soil in PR China. The taxonomic position of this strain was investigated using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain ZZ-4T was a member of the genus Emticicia and showed the highest sequence similarity to Emticicia fontis IMCC1731T (98.0 %) and Emticicia ginsengisoli Gsoil 085T (97.2 %), and lower (<97 %) sequence similarity to other known Emticicia species. Chemotaxonomic analysis revealed that strain ZZ-4T possessed menaquinone MK-7 as the major isoprenoid quinone; and iso-C15 : 0, summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7), iso-C17 : 0 3-OH and C16 : 1ω5c were the predominant fatty acids. Strain ZZ-4T showed low DNA-DNA relatedness with E. fontis IMCC1731T (39.8±3.1 %) and E. ginsengisoli Gsoil 085T (44.51±1.5 %). The DNA G+C content was 38.3 mol%. Based on the phylogenetic and phenotypic characteristics, chemotaxonomic data and DNA-DNA hybridization results, strain ZZ-4T is considered to represent a novel species of the genus Emticicia, for which the name Emticicia soli sp. nov. is proposed. The type strain is ZZ-4T (=KCTC 52344T=CCTCC AB 2016137T).

Terrimonas suqianensis sp. nov., isolated from a tetrabromobisphenol A-contaminated soil.

Strain C3-5T, a Gram-negative, asporogenous, rod-shaped bacterium, was isolated from a tetrabromobisphenol A contaminated soil. Growth was observed at 10-37 °C (optimum 30 °C) and at pH 5.5-9.5 (optimum pH 7.0). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain C3-5T is a member of the genus Terrimonas and exhibits high sequence similarities with Terrimonas pekingensis QHT (99.0%) and Terrimonas rhizosphaerae CR94T (97.3%), and exhibits low (<97%) sequence similarities with other known Terrimonas species. Chemotaxonomic analysis revealed that strain C3-5T possesses menaquinone-7 (MK-7) as the major isoprenoid quinone and iso-C15:0, iso-C15:1 G, iso-C17:0 3-OH and summed feature 3 (C16:1 ω6c and/or C16:1 ω7c) as the major (>5% of total) fatty acids. The polar lipids were determined to be a lipid, glycolipid, phospholipid, phosphoaminolipid and phosphatidylethanolamine. The DNA G+C content was found to be 42.6 mol%. The DNA-DNA relatedness values with the closely related strains T. pekingensis QHT and T. rhizosphaerae CR94T were 25.2 and 48.5%, respectively. Based on phylogenetic analysis, phenotypic characteristics and chemotaxonomic data, strain C3-5T is considered to represent a novel species of the genus Terrimonas, for which the name Terrimonas suqianensis sp. nov. is proposed. The type strain is C3-5T (= CCTCC AB 2017042T = KCTC 52676T).

Association between CLPTM1L-TERT rs401681 polymorphism and risk of pancreatic cancer: a meta-analysis.

Telomere biology plays a critical and complex role in the initiation and progression of cancer. Several recent studies have provided evidence that rs401681 polymorphisms in intronic region of cleft lip and palate trans-membrane 1-like (CLPTM1L) gene sequence are associated with pancreatic cancer (PC) development, but a comprehensive synopsis is not available. We performed a meta-analysis of 6 case-control studies that included 8,253 pancreatic cancer cases and 37,646 case-free controls. We assessed the strength of the association, using odds ratios (ORs) with 95 % confidence intervals (CIs). Overall, this meta-analysis showed that rs401681 allele T was associated with a significantly increased PC risk (OR = 1.17, 95 % CI = 1.12-1.22, P heterpgeneity = 0.596 and I (2) = 0). Similarly, in the subgroup analysis by ethnicity, a significantly increased risk was found among Asians (OR = 1.15, 95 % CI = 1.07-1.24, P heterpgeneity = 0.297 and I (2) = 8.0 %) and among Caucasian (OR = 1.13, 95 % CI = 1.02-1.26, P heterpgeneity = 0.385 and I (2) = 0). No publication bias was found in the present study. This meta-analysis suggests that T allele of CLPTM1L-telomerase reverse transcriptase rs401681 polymorphism is associated with an increased PC risk, especially among Chinese. Further large and well-designed studies are needed to confirm this association.

[Role of Smad signaling in transcription of Smad7 gene mediated by TGF-beta1 in odontoblast cell line MDPC-23].

PURPOSE: To investigate the role of Smad signaling in transcription of Smad7 gene mediated by TGF-beta1 in odontoblast cell line MDPC-23, and to explore the molecular mechanism of Smad7 gene expression mediated by TGF-beta1 at the transcriptional level. METHODS: Smad function and its role in transcription of Smad7 were investigated in cotransfection experiments using Smad7 promoter-luciferase reporter construct containing the sequence between -408 bp and +112 bp of mouse Smad7 gene. The data were analysed by one-way ANOVA. RESULTS: When the Smad7 promoter-luciferase reporter gene construct was expressed in MDPC-23 cells, its transcriptional activity was significantly induced by TGF-beta1 treatment, whereas not by BMP-2 treatment. Overexpression of Smad1, 2, 4, or 5 had no effect on transcriptional activity of Smad7 promoter. Overexpression of Smad3 markedly promoted transcriptional activity of Smad7 promoter, whereas co-transfection of Smad3 and Smad4 doubled the effect of Smad3. Overexpression of Smad3 dominant negative mutant or Smad3 antisense cDNA (AS-Smad3) significantly inhibited transcriptional activity of Smad7 promoter induced by TGF-beta1. CONCLUSION: TGF-beta1 regulated transcription of Smad7 gene through association of Smad3 and Smad4 in MDPC-23 cells.

[Cloning, expression and antitumor effect of mouse costimulatory molecule 4-1BBL].

AIM: To clone mouse 4-1BBL gene, construct its eukaryotic expression vector, and evaluate antitumor activity of the expression product. METHODS: RT-PCR was used to amplify mouse 4-1BBL gene from total RNA of C57BL/6 splenocytes stimulated by PHA. Then m4-1BBL cDNA was subcloned into eukaryotic expression vector pcDNA3.1(+) and transfected into mouse hepatocellular carcinoma cell line Hepa1-6. The expression of m4-1BBL in transfected cells was detected by RT-PCR, indirect immunofluorescence staining, and flow cytometry. Non-adherent splenocytes from non-immunized C57BL/6 mice were incubated with mitomycin-treated non-transfected Hepa1-6(Hepa1-6-wt) or transfected Hepa1-6 cells (Hepal-6-m4-1BBL), respectively. Then the lymphocytes were tested for cytotoxic activity to Hepa1-6-wt cells. RESULTS: The Hepa1-6 cells transfected by pcDNA3.1(+)-m4-1BBL could efficiently express m4-1BBL. As compared with Hepa1-6-wt cells,Hepa1-6-m4-1BBL cells could induce more efficiently cytotoxic activity of lymphocytes to Hepa1-6-wt cells (P<0.01). CONCLUSION: The expression of m4-1BBL by tumor cells is effective in inducing antitumor immune response.

[Antitumor immune responses induced by gene transfer of 4-1BBL into hepatocellular carcinoma Hepa1-6 in vitro].

OBJECTIVE: To study the cytotoxic activity against tumor cells and cytokines production of spleen cells induced in vitro by murine 4-1BBL gene transfected Hepa1-6. METHODS: The eukaryotic expression vector pCDNA3.1(+)-m4-1BBL was transfected into murine hepatocellular carcinoma cell line Hepa1-6 by Liposomes. Then the transfected cells were selected in medium containing G418 (400 - 800 micro g/ml) and termed as Hepa1-6-m4-1BBL. The TCV-m4-1BBL was obtained by treating them with mitomycin (MMC). Cocultivation TCV with syngeneic murine spleen cells, then the lymphocytes were tested for cytotoxic activity against Hepa1-6-wt cells and the supernatants were harvested for detecting the cytokines (IL-2, TNF-alpha and GM-CSF). RESULTS: Hepa1-6-m4-1BBL cells expressed 4-1BBL protein with highest cell surface level. The 4-1BBL mRNA could still be detected in the cells when cultured 48 h after treated with MMC (80 mg/L). Comparing with TCV-Hepa1-6, the tumor cell vaccine derived from Hepa1-6-m4-1BBL (TCV-m4-1BBL) could induce a more efficient cytotoxic activity of syngeneic murine lymphocyte against its parental tumor cell Hepa1-6 (P < 0.05), but not against non-parental tumor cell H22 and NIH3T3. Higher levels of IL-2, TNF-alpha and GM-CSF were released by the splencytes after stimulated by TCV-m4-1BBL. CONCLUSIONS: These results suggest the expression of m4-1BBL by tumor cells is effective in inducing antitumor immune responses.