Reclassification of Sabulilitoribacter multivorans and Sabulilitoribacter arenilitoris as Flaviramulus multivorans comb. nov. and Wocania arenilitoris comb. nov., respectively, based on genome sequence analysis.

Rapid advancements in DNA sequencing technologies are providing new approaches for bacterial taxonomy. The genus Sabulilitoribacter is a member of the family Flavobacteriaceae, which consists of more than 150 genera. In this study, genome sequence analysis was conducted to revisit the taxonomic status of Sabulilitoribacter arenilitoris and Sabulilitoribacter multivorans, the only two species of this genus. Genome sequence based phylogeny analysis showed that the genus Sabulilitoribacter was non-monophyletic: S. multivorans, the type species of genus Sabulilitoribacter, was clustered with the type species of the genus Flaviramulus, whereas S. arenilitoris formed a robust cluster with the only two species of the genus Wocania. The values of average amino acid identity, genome-wide average nucleotide identity, alignment fractions and some phenotypic features showed that S. multivorans was more closely related with the type species of the genus Flaviramulus than with S. arenilitoris, and S. arenilitoris was more closely related with the only two species of the genus Wocania than with S. multivorans. Based on these results, we consequently propose that S. multivorans and S. arenilitoris should be reclassified as Flaviramulus multivorans comb. nov. and Wocania arenilitoris comb. nov. respectively.

EB virus-positive tumors are inhibited by rBCG expressing hGM-CSF and LMP2A.

For the development of safe and effective EBV (Epstein-Barr virus) vaccines, the Ag85A signal peptide from M. tuberculosis H37Rv was used to construct a recombinant secretory BCG (Bacillus Chalmette-Guérin) plasmid. The Ag85A gene, fused to the EBV LMP2A (latent membrane protein) and hGM-CSF (human granulocyte/macrophage colony-stimulating factor) genes, was inserted into the pMV261 vector (secretory BCG plasmid). The expression levels of the hGM-CSF and LMP2A proteins in rBCG (recombinant BCG) were measured by Western blot analysis. Humoral immunity, cellular immunity, and antitumor effects were determined by a series of experiments. The recombinant pMVGCA plasmid effectively expressed GCA (hGM-CSF and LMP2A fusion protein) in BCG after transformation, and the rBCG proteins were recognized by antibodies against hGM-CSF and LMP2A. Six weeks after immunization, the maximum dose of rBCG resulted in antibody titers of 1:19,800 (hGM-CSF antibody) and 1:21,800 (LMP2A antibody). When the effector:target ratio was 40:1, specific lysis was maximal and approximately two times stronger than that in mice immunized with the control. Tumorigenicity was lower in the rBCG treatment group, with a tumor inhibition rate of 0.81 ± 0.09 compared with the control groups. EB virus-positive tumors are inhibited by rBCG expressing an hGM-CSF and LMP2A fusion protein.

Litorilituus lipolyticus sp. nov., isolated from intertidal sand of the Yellow Sea in China, and emended description of Colwellia asteriadis.

A Gram-stain negative, rod-shaped, aerobic, oxidase-positive and catalase-weakly positive bacterial strain with polar or subpolar flagellum, designated RZ04T, was isolated from an intertidal sand sample collected from a coastal area of the Yellow Sea, China. The organism was observed to grow optimally at 25 °C and pH 6.5-7.0 with 2% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain RZ04T was closely related to Colwellia asteriadis (similarity 96.9%) and Litorilituus sediminis (similarity 96.8%), and 94.4-96.4% sequence similarities to other type strains of species of the genera belonged to the family Colwelliaceae. The dominant fatty acids of strain RZ04T were determined to be C17:1ω8c, C15:1ω8c, C16:0 and summed feature 3 (C16:1ω6c and/or C16:1ω7c), and the predominant isoprenoid quinone was determined to be quinone 8 (Q-8). Phosphatidylethanolamine, phosphatidylglycerol, an unidentified aminophospholipid and four unidentified lipids were determined to be the major constituents of the polar lipids. The genome of strain RZ04T is 4.14 Mbp with a G + C content of 37.4 mol%. A total of 3631 genes are predicted, with 3531 protein-coding genes, 75 RNA genes and 25 pseudogenes. Based on phenotypic, genotypic and phylogenetic analysis, strain RZ04T is considered to represent a novel species in the genus Litorilituus, for which the name Litorilituus lipolyticus is proposed. The type strain is RZ04T (= MCCC 1K03616T = KCTC 62835T). An emended description of Colwellia asteriadis is also provided.

Flavivirga rizhaonensis sp. nov., a marine bacterium isolated from intertidal sand.

A bacterial strain designated RZ03T was isolated from an intertidal sand sample from the Yellow Sea in China and characterised using a polyphasic taxonomic approach. Cells of strain RZ03T were observed to be Gram-stain negative, aerobic, and oxidase and catalase positive rods showing gliding motility and forming yellow colonies. Growth was found to occur at 7-30 °C (optimum, 25 °C), at pH 5.5-9.5 (optimum, pH 6.5-7.0) and with 0.5-5% NaCl (optimum, 1.5-2%). Phylogenetic analysis based on 16S rRNA gene sequences indicates that strain RZ03T clusters within members of the genus Flavivirga of the family Flavobacteriaceae and is closely related to the type strains Flavivirga amylovorans JCM 17112T and Flavivirga jejuensis JCM 17113T (97.9% and 97.5% similarity, respectively). The predominant cellular fatty acids are iso-C15:0, iso-C15:1 G, iso-C17:0 3-OH and iso-C15:0 3-OH and the major respiratory quinone is MK-6. Polar lipids include phosphatidylethanolamine, three unidentified aminolipids, an unidentified phospholipid and four unidentified lipids. The genome of strain RZ03T is 4.88 Mbp with a G+C content of 32.2 mol%. A total of 4152 genes are predicted, with 4052 protein-coding genes, 51 RNA genes and 49 pseudogenes. This polyphasic study suggests that strain RZ03T represents a novel species in the genus Flavivirga, for which the name Flavivirga rizhaonensis is proposed. The type strain is RZ03T(= KCTC 62833T = MCCC 1K03615T).

AXIN2 is Associated With Papillary Thyroid Carcinoma.

BACKGROUND: Findings of recent studies have demonstrated a rapid increase of the incidence of papillary thyroid carcinoma (PTC), which accounts for nearly 80% of thyroid cancers. OBJECTIVES: The aim of this study was to explore the association between AXIN2 gene polymorphism and papillary thyroid carcinoma (PTC). PATIENTS AND METHODS: 106 blood samples (56 PTC patients and 50 healthy controls) were drawn from China-Japan Union Hospital in Jilin province, China, during October 2010 to March 2011. A case-control study was designed to examine the association between AXIN2 and PTC. Seven tag single nucleotide polymorphisms (tag SNPs) in AXIN2 were selected and genotyped. Frequencies of different genotypes and alleles were analyzed between the patients and the controls, using the R × C column contingency table χ(2) test. The possible association of haplotypes constructed by the combined effects of two or more loci with PTC was analyzed through the UNPHASED 3.1.4 program. RESULTS: Rs11655966, rs3923086 and rs7591 of AXIN2 showed significant associations with PTC (P < 0.05). The result of haplotypes analysis showed that rs11655966-rs3923086-rs4791169 had statistically significant differences between the two groups (P < 0.05). CONCLUSIONS: Together with the functions of the target genes, we further elucidated that AXIN2 is associated with papillary thyroid carcinoma in the Chinese Han population.

Biocontrol of Gray Mold Decay in Pear by Bacillus amyloliquefaciens Strain BA3 and its Effect on Postharvest Quality Parameters.

The economic losses caused by postharvest fruits diseases have attracted global attention. Traditional chemical fungicide could not meet the need of humans. In recent years, microbial agent which has begun to take the place of chemical fungicide comes into people's vision. The aim of this paper was to investigate the potential of Bacillus amyloliquefaciens strain BA3 for its biocontrol capability on gray mold decay of pears and its effect on postharvest quality of pears. Compared with other treatments, the inhibition effect on gray mold of washed cell suspension of B. amyloliquefaciens was the best. Consequently it was utilized in subsequent experiments. Spore germination and germ tube length of Botrytis cinerea was 18.72% and 12.85 µm treated with BA3, while the control group was 62.88% and 30.44 µm. We confirmed that increase of the concentration of B. amyloliquefaciens, improved the efficacy of BA3 in controlling gray mold decay of pears. Colonization variation of BA3 in wounds of pears was recorded. To begin with, the populations of B. amyloliquefaciens increased rapidly and remained stable. On the fourth day, there was a declining trend , after that the population increased to 4 × 105 CFU/wound and remained stable. BA3 had no significant effect on mass loss, titratable acidity, firmness and total soluble solids of pears that were stored at 25°C for 7 days comparing with control group. However, the effect of B. amyloliquefaciens on ascorbic acid was significantly higher than that of the control group. Our study indicates that B. amyloliquefaciens has a potential as postharvest biocontrol agent on pears.

Biocontrol of Gray Mold Decay in Pear by Bacillus amyloliquefaciens Strain BA3 and its Effect on Postharvest Quality Parameters.

The economic losses caused by postharvest fruits diseases have attracted global attention. Traditional chemical fungicide could not meet the need of humans. In recent years, microbial agent which has begun to take the place of chemical fungicide comes into people's vision. The aim of this paper was to investigate the potential of Bacillus amyloliquefaciens strain BA3 for its biocontrol capability on gray mold decay of pears and its effect on postharvest quality of pears. Compared with other treatments, the inhibition effect on gray mold of washed cell suspension of B. amyloliquefaciens was the best. Consequently it was utilized in subsequent experiments. Spore germination and germ tube length of Botrytis cinerea was 18.72% and 12.85 µm treated with BA3, while the control group was 62.88% and 30.44 µm. We confirmed that increase of the concentration of B. amyloliquefaciens, improved the efficacy of BA3 in controlling gray mold decay of pears. Colonization variation of BA3 in wounds of pears was recorded. To begin with, the populations of B. amyloliquefaciens increased rapidly and remained stable. On the fourth day, there was a declining trend , after that the population increased to 4 × 10(5) CFU/wound and remained stable. BA3 had no significant effect on mass loss, titratable acidity, firmness and total soluble solids of pears that were stored at 25°C for 7 days comparing with control group. However, the effect of B. amyloliquefaciens on ascorbic acid was significantly higher than that of the control group. Our study indicates that B. amyloliquefaciens has a potential as postharvest biocontrol agent on pears.

Development of a MAb-based immunoassay for the simultaneous determination of O,O-diethyl and O,O-dimethyl organophosphorus pesticides in vegetable and fruit samples pretreated with QuEChERS.

To develop a broad-specificity immunoassay for organophosphorus pesticides (OPs), a broad-specificity monoclonal antibody (MAb) for OPs against a generic hapten, O,O-diethyl O-(3-carboxyphenyl) phosphorothioate with the carboxy group in the meta position of the benzene ring, was produced. Eight haptens were prepared and covalently attached to ovalbumin (OVA) for use as coating antigens, and the optimum coating antigen was selected. Then, a sensitive and broadly class selective competitive indirect enzyme-linked immunosorbent assay (ciELISA) based on the MAb and the optimum coating antigen (hapten H-OVA, possessing an O,O-dimethyl generic structure and linked through a linear spacer arm) was developed and optimized. The MAb developed in this study showed quite different cross-reactivity and selectivity compared to previously produced anti-OPs broad-specificity MAbs. Specifically, the MAb showed high and uniform sensitivity to seven O,O-diethyl OPs and six O,O-dimethyl OPs. With the optimum ciELISA, the IC50 values of the 13 OPs were determined as 23.1∼151.2 ng mL(-1). The average IC50 and coefficient of variation (CV) for the IC50 values of the 13 OPs were 74.6 ng mL(-1) and 33.9%, respectively. For the recovery study, a QuEChERS approach based on dispersive solid-phase extraction (d-SPE) was implemented to decrease the matrix effects of vegetable and fruit samples. The recoveries of six representative OPs from the spiked samples ranged from 89.4 to 135.5%; the CV ranged from 3.5 to 15.7%. The ciELISA was also applied to real samples, followed by confirmation with gas chromatography-tandem mass spectrometry (GC-MS/MS) analysis. The results demonstrated that the ciELISA is suitable for monitoring OP contamination in vegetable and fruit samples.

[Association of BRAF(V) 600E mutation with clinical pathological features in papillary thyroid carcinoma: meta-analysis].

OBJECTIVE: To evaluate the association between BRAF(V) 600E mutation and pathological features in papillary thyroid carcinoma (PTC). METHODS: The raleted studies were searched through Medline, PubMed, and Web of Science, and meta-analysis was used to calculate the OR and 95%CI of the study results. RESULTS: A total of 52 studies including 12 029 PTC patients was identified. The prevalence of BRAF(V) 600E mutation in PTC was 57.85%. There was a closed association between the BRAF(V) 600E mutation and pathological features, including advanced TNM stage (OR = 1.89, 95%CI 1.58-2.27), lymph node metastasis (OR = 1.74, 95%CI 1.42-2.14), multifocality (OR = 1.25, 95%CI 1.07-1.46), and recurrence (OR = 2.26, 95%CI 1.25-4.09) of PTC. For Asians with PTC, the association between the BRAF(V) 600E mutation and pathological features was indicated for advanced TNM stage (OR = 1.56, 95%CI 1.24-1.96) and lymph node metastasis (OR = 1.57, 95%CI 1.25-1.99). For Europeans with PTC, the association between the BRAF(V) 600E mutation and pathological features was indicated for advanced TNM stage (OR = 2.63, 95%CI 2.10-3.30), lymph node metastasis(OR = 1.97, 95%CI 1.19-3.25), and multifocality (OR = 1.35, 95%CI 1.01-1.80). CONCLUSION: There were associations between the BRAF(V) 600E utation and advanced TNM stage, lymph node metastasis, multifocality, and recurrence of PTC.

Body mass index and risk of gastric cancer: a meta-analysis.

OBJECTIVE: Overweight and obesity, indicated as increased body mass index, are associated with the risk of some cancers. We carried out a meta-analysis on published cohort and case-control studies to assess the strength of association between body mass index and gastric cancer. METHODS: Relevant studies were identified through PubMed, Web of Science and Medline electronic databases. Adjusted relative risks (odds ratios) with 95% confidence interval were used to assess the strength of association between body mass index and gastric cancer. RESULTS: Sixteen eligible studies were included in this meta-analysis. Overall, obesity (body mass index ≥ 30 kg/m(2)) was associated with an increased risk of gastric cancer (odds ratio = 1.13, 95% confidence interval = 1.03-1.24) compared with normal weight (body mass index = 18.5 to <25 kg/m(2)), while overweight (body mass index = 18.5 to <30 kg/m(2)) showed no association (odds ratio = 1.04, 95% confidence interval = 0.96-1.12). Specifically, a stratified analysis showed there were associations between obesity and the increased risk of gastric cancer for males (odds ratio = 1.27, 95% confidence interval = 1.09-1.48), non-Asians (odds ratio = 1.14, 95% confidence interval = 1.02-1.28) and both cohort studies (odds ratio = 1.10, 95% confidence interval = 1.00-1.22) and case-control studies (odds ratio = 1.29, 95% confidence interval = 1.03-1.60). Both overweight (odds ratio = 1.22, 95% confidence interval = 1.05-1.42) and obesity (odds ratio = 1.61, 95% confidence interval = 1.15-2.24) were associated with the increased risk of gastric cardia cancer. CONCLUSIONS: The results indicated that obesity was associated with the risk of gastric cancer, especially for males and among non-Asians. Both overweight and obesity were associated with the risk of gastric cardia cancer.

Magnetic fields exposure and childhood leukemia risk: a meta-analysis based on 11,699 cases and 13,194 controls.

OBJECTIVE: To observe the association between childhood leukemia and magnetic field exposure. METHODS: The literature was searched by PubMed, ProQuest, Web of Science (SCI) and Medline databases during 1997-2013. Heterogeneity in several studies was weighted by I-squared value. Publication bias was tested by funnel plot and Egger's test. Odds ratio (OR) and 95% CI were used to evaluate the association strength. The statistical analyses in present study were carried out by STATA software package (version 12.0, College Station, TX). RESULTS: A total of 11,699 cases and 13,194 controls in 9 studies were stratified by different exposure cut-off points. On condition of the reference <0.1 µT, statistical association between magnetic field intensity ≥0.4 µT and childhood leukemia was exhibited (for total leukemia: OR = 1.57, 95% CI = 1.03-2.40; for acute lymphocytic leukemia: OR=2.43, 95% CI = 1.30-4.55). On condition of the reference level of <0.2 µT, the positive association between magnetic field intensity ≥0.2 µT and childhood leukemia was found (OR = 1.31, 95% CI = 1.06-1.61). CONCLUSIONS: The result in this meta-analysis indicated that magnetic field exposure level may be associated with childhood leukemia.

The association between BRAF (V600E) mutation and pathological features in PTC.

The BRAF (V600E) mutation is a common genetic alteration in papillary thyroid carcinoma (PTC) and is associated with some pathological features. The association has been widely reported, but results were inconclusive. In this study a meta-analysis was done to explore the association between BRAF(V600E) mutation and pathological features in papillary thyroid carcinoma. Medline, PubMed and Web of Science were searched. A total of 69 studies that included 14,170 PTC patients were identified. The outcomes were from 2004 to October 2013. STATA12.0 software package was used to analyze the data. The result was assessed based on pooled odds ratios (ORs) with 95 % confidence intervals (CIs). The results showed that the BRAF (V600E) mutation was associated with extra-thyroidal extension (OR = 2.09, 95 % CI = 1.69-2.58), advanced TNM stage (OR = 1.90, 95 % CI = 1.62-2.22), lymph node metastasis (OR = 1.68, 95 % CI = 1.41-2.01), multifocality (OR = 1.22, 95 % CI = 1.06-1.40), and recurrence (OR = 2.50, 95 % CI = 1.73-3.59). The meta-analysis suggested the potential roles of BRAF (V600E) mutation in pathological features. BRAF (V600E) might provide prognostic and diagnostic information for papillary thyroid carcinoma.

[Effect of exposure to extremely low-frequency electromagnetic fields on liver function of workers].

OBJECTIVE: To examine the effect of exposure to extremely low-frequency electromagnetic fields (ELF EMFs) on the liver function of workers. METHODS: The workers in a factory were selected as subjects, and the recent physical examination data of these workers were collected. The workers aged 20∼40 years and with more than 2 years' working experience were included for analysis; considering the intensity of electromagnetic field, the workers exposed to less electromagnetic radiation were assigned to exposure I group (n = 123), those exposed to more electromagnetic radiation to exposure II group (n = 229), and those not exposed to electromagnetic radiation to control group (n = 212). There were no significant differences in sex, age, height, and body weight between the three groups (P > 0.05). Physical examination, including measurements of direct bilirubin (DBil), alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (GGT), and albumin, was performed in a health examination center. The intensity of electromagnetic field was measured by EFA-300 power frequency electromagnetic field analyzer, and the intensity of noise by AWA5610D integrating sound level meter. RESULTS: The intensities of electric field and the magnetic field in exposure II group were significantly higher than those in the exposure I group. The levels of ALT, ALP, AST, GGT and albumin in exposure II group were significantly higher than those in exposure I group and control group. However, the level of direct bilirubin in exposure II group was significantly lower than that in exposure I group and control group. CONCLUSION: Occupational exposure to ELF EMFs may affect human liver function.

Effects of extremely low frequency electromagnetic field on the health of workers in automotive industry.

OBJECTIVE: To observe the effects of extremely low frequency electromagnetic fields (ELF-EMFs) in automotive industry on occupational workers. METHODS: A total of 704 workers were investigated, and 374 workers were chosen and divided into two groups (control group and exposure group) according to the inclusive criteria, namely male with age 20-40 years old and ≥ 2 years of exposure. The intensities of ELF-EMFs and noise were detected with EFA-300 Field Analyzer (Narda company, Pfullingen, Germany) and AWA5610D integrating sound level meter (Hangzhou Aihua Instruments Co., Ltd, Hangzhou, China), respectively. Survey data were collected by questionnaire, and the physical check-up was done in hospital. All the data were input into SPSS17.0 software (SPSS Inc, Chicago, USA), and the appropriate statistic analyses were carried out. RESULTS: The intensity of EMFs in exposure group was significantly higher than that in control group (p < 0.05), while the noise in two workplaces showed no difference (p>0.05). The survey data collected by questionnaires showed that the symptoms of loss of hair in exposure group were significantly different as compared with that in control group (p < 0.05). The check-up parameters of cardiovascular, liver and hematology system showed significant differences between the two groups (p < 0.05). CONCLUSIONS: Survey and check-up data suggest that exposure to ELF-EMFs might have effects on the nervous, cardiovascular, liver, and hematology system of workers.

Expression profiles of microRNAs and their target genes in papillary thyroid carcinoma.

The incidence of thyroid cancer has recently experienced a rapid increase in China, and papillary thyroid carcinoma (PTC) accounts for nearly 80% of human thyroid cancers. In the present study, the differential expression of microRNAs (miRNAs) and their target genes were identified in order to analyze the potential roles of miRNAs as biomarkers and in papillary thyroid carcinogenesis. One hundred and twenty-six PTC samples were collected from patients at the China-Japan Union Hospital, China, and the gene/miRNA expression profiles were examined with Illumina BeadChips and verified by real­time RT-PCR. Gene Ontology (GO) categories were determined, and pathway analysis was carried out using KEGG. miRNA target genes were predicted by implementing three computational analysis programs: TargetScanS, DIANA-microT and PicTar. Two hundred and forty-eight miRNAs and 3,631 genes were found to be significantly deregulated (gene, P<0.05; miRNA, P<0.01) in PTC tissues when compared with their matching normal thyroid tissues. hsa-miR-206 (target gene, MET), hsa-miR-299-3p (target gene, ITGAV), hsa-miR-101 (target gene, ITGA3), hsa-miR-103 (target gene, ITGA2), hsa-miR­222 (target genes, KIT and AXIN2), hsa-miR-15a (target genes, AXIN2 and FOXO1) and hsa-mir-221 (target gene, KIT) were identified. Together with the functions of the target genes, we further elucidated the role of miRNAs in papillary thyroid carcinogenesis and suggest the use of miRNAs as biomarkers for early diagnosis. Our findings provide the basis for future studies in the field of miRNA-based cancer therapy.

[Effects of extremely low frequency electromagnetic radiation on cardiovascular system of workers].

OBJECTIVE: To observe the exposure levels of extremely low frequency electromagnetic fields in workplaces and to analyze the effects of extremely low frequency electromagnetic radiation on cardiovascular system of occupationally exposed people. METHOD: Intensity of electromagnetic fields in two workplaces (control and exposure groups) was detected with EFA-300 frequency electromagnetic field strength tester, and intensity of the noise was detected with AWA5610D integral sound level. The information of health physical indicators of 188 controls and 642 occupationally exposed workers was collected. Data were analyzed by SPSS17.0 statistic software. RESULTS: The intensity of electric fields and the magnetic fields in exposure groups was significantly higher than that in control group (P < 0.05), but there was no significant difference of noise between two workplaces (P > 0.05). The results of physical examination showed that the abnormal rates of HCY, ALT, AST, GGT, ECG in the exposure group were significantly higher than those in control group (P < 0.05). There were no differences of sex, age, height, weight between two groups (P > 0.05). CONCLUSION: Exposure to extremely low frequency electromagnetic radiation may have some effects on the cardiovascular system of workers.