Methylobacterium crusticola sp. nov., isolated from biological soil crusts.

A pink-pigmented, Gram-negative, rod-shaped, obligate aerobic bacterial strain, MIMD6T, was isolated from biological soil crusts in PR China. Cells grew at 20-37 °C (optimum, 30 °C), at pH 6-8 (optimum, pH 7) and with 0-1 % (w/v) NaCl (optimum, 0 %). Strain MIMD6T could use methanol or formate as a sole carbon source to grow, and carried methanol dehydrogenase genes mxaF and xoxF, supporting its methylotrophic metabolism. The respiratory quinone was ubiquinone Q-10, the major fatty acids were C18 : 1ω7c (87.3 %), and the major polar lipids were diphosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, one unknown aminolipid and one unidentified glycolipid. The results of phylogenetic analyses based on the sequences of the 16S rRNA gene, seven housekeeping genes (dnaK, recA, rimO, rpIK, rpmG, rpsR and rpoB) and methanol dehydrogenase genes indicated that strain MIMD6T formed a phylogenetic linage with members of the genus Methylobacterium. Strain MIMD6T was most closely related to Methylobacterium isbiliense DSM 17168T and Methylobacterium nodulans LMG 21967T with 16S rRNA gene sequence similarities of 95.7 and 95.2 %, respectively. The genomic DNA G+C content calculated via draft genome sequencing was 73.0 mol%. The average nucleotide identity and digital DNA-DNA hybridization values between strain MIMD6T and the type strains of other Methylobacterium species were 70.7-82.0 and 24.6-30.0 %, respectively. Based on phenotypic, chemotaxonomic and phylogenetic characteristics, strain MIMD6T represents a novel species of the genus Methylobacterium, for which the name Methylobacterium crusticola sp. nov. is proposed. The type strain is MIMD6T (=KCTC 52305T=MCCC 1K01311T).

[Response of Nitrification Potential and Ammonia Oxidation Microbial Community in Purple Soils to the Application of Biochar Combined with Chemical Fertilizer and Manure].

The effects of manure and chemical fertilizer combined with biochar on nitrification potential and ammonia oxidation microbial change characteristics of purple soils were studied to explore the effects of fertilization measures and soil environmental factors on nitrification potential and ammonia oxidation microbial change characteristics. In this study, purple soil was taken as the research object, and five treatments were set up:no fertilizer(CK), chemical fertilizer(F), manure(P), chemical fertilizer plus biochar(FP), and manure plus biochar(PP). PCR and T-RFLP methods were used to study the characteristics of soil AOA and AOB communities, and soil nitrification potential and environmental factors were measured at the same time to determine the effect of manure combined with biochar on the nitrification potential of purple soil. The results showed that:① compared with that in the CK treatment, the FP and PP treatments significantly increased soil nitrification potential(P<0.05). ② Compared with that in the CK treatment, the F, P, FP, and PP treatments significantly increased the number of copies of the AOA amoA gene in the soil by 78.17%-162.22%(P<0.05), and the F, FP, and PP treatments significantly increased the number of copies of the AOB amoA gene by 21.56%-78.32%(P<0.05). ③ Compared with that in the CK treatment, the PP treatment significantly improved the Shannon, richness, and evenness indices of the soil AOA community(P<0.05), and the combination of biochar(FP and PP treatments) could change the community structure of the soil AOB. ④ The chemical fertilizer and manure affected the AOA and AOB community structure by changing soil properties such as pH, TP, AP, C/N, SOM, NO3--N, and NH4+-N. ⑤ Stepwise regression analysis further showed that AOB amoA gene abundance was the main factor affecting soil nitrification potential. Therefore, under the condition that chemical fertilizer and manure were applied together with biochar, the AOB community was the main driver of the ammonia oxidation process in acidic purple soil, and fertilization affected the ammonia oxidation process by regulating C/N and NO3--N.

[Effect of Adding Granular Activated Carbon (GAC)/Manganese Dioxide (MnO2) for the Anaerobic Digestion of Waste Activated Sludge].

The addition of conductive materials or metal oxide nanoparticles to an anaerobic system is an attractive strategy to enhance anaerobic digestion. The effect of granular activated carbon (GAC) and/or manganese dioxide (MnO2) on waste activated sludge (WAS) anaerobic digestion was investigated by batch experiments. The experiments were provided in control, GAC, MnO2, and GAC/MnO2 groups, which were named R0, R1, R2, and R3, respectively. The sludge characteristics, microbial activity, and microbial community structure were systematically investigated. Results showed that CH4 yield rate was evidently increased by 68.18% and 51.35% in R1 and R3, respectively, whereas the cumulative CH4 production decreased by 21.25% in R2, compared with R0. Moreover, the fermentation process could be promoted with the addition of GAC and/or MnO2. The phosphate precipitation generated by Mn2+ and phosphate released from WAS blocked the anaerobic metabolic channel and then decreased the production of CH4 in R2. The increase in CH4 yield rate in R3 was mainly attributed to the conductivity and adsorption of GAC and the catalysis of MnO2. Additionally, the microbial activity could be promoted with the addition of GAC, MnO2, and GAC/MnO2 in anaerobic digestion. Microbial community structure analysis showed that the abundance of the Methanobacterium and Methanosaeta increased with the addition of GAC and MnO2, which could enhance the interspecies electron transfer between fermenting bacteria and methanogens and boost fermentation and CH4 production.

The efficacy of postoperative radiotherapy in localized primary soft tissue sarcoma treated with conservative surgery.

BACKGROUND: To evaluate the efficacy of postoperative radiotherapy (RT) on local failure-free survival (LFFS), distant metastasis-free survival (DMFS) and overall survival (OS) in patients with localized primary soft tissue sarcoma (STS) and to identify prognostic factors. METHODS AND MATERIALS: Between January 2000 and July 2010, 220 consecutive patients with localized primary STS, who received conservative surgery with or without postoperative RT, were enrolled in the study. Survival curves were constructed by the Kaplan-Meier method and log-rank test was used to assess statistical significance. Multivariate analysis was applied to identify the prognostic factors. RESULTS: After a median follow-up of 68 months (range, 5-127 months), the 5-year LFFS, DMFS and OS were 70.0, 78.2 and 71.2 %, respectively. Tumor size, histological subtypes, margin status and postoperative RT were independent predictors for OS. Postoperative RT was associated with a significant reduced local recurrence risk versus surgery alone (hazard ratio [HR] = 0.408, 95 % confidence interval [CI] 0.235-0.707, P = 0.001), with 5-year LFFS of 81.1 and 63.6 %, respectively (log-rank, P = 0.004). The log-rank test showed that postoperative RT had a tendency of improving OS compared with surgery alone, with 5-year OS of 74.8 and 65.0 %, respectively (P = 0.089). Multivariate analysis demonstrated that postoperative RT significantly reduced mortality rate compared with surgery alone (HR = 0.512, 95 % CI 0.296-0.886, p = 0.017), especially in patients with liposarcoma (p = 0.034). CONCLUSION: Postoperative radiotherapy reduce both local recurrence and STS mortality in patients with localized primary STS. The efficacy of RT on survival warrants further prospective study.

Avian leukosis virus subgroup J triggers caspase-1-mediated inflammatory response in chick livers.

Many pathogens trigger caspase-1-mediated innate immune responses. Avian leukosis virus subgroup J (ALV-J) causes serious immunosuppression and diverse tumors in chicks. The caspase-1 inflammasome mechanism of response to ALV-J invading remains unclear. Here we investigated the expression of caspase-1, the inflammasome adaptor NLRP3, IL-1ß and IL-18 in response to ALV-J infection in the liver of chick. We found caspase-1 mRNA expression was elevated at 5 dpi and peaked at 7 dpi in ALV-J infected animals. Corresponding to this, the expressions of NLRP3 and proinflammatory cytokines IL-1ß and IL-18 were significantly increased at 5 or 7 dpi. In addition, caspase-1 protein expression and inflammatory cell infiltration were induced after virus infection. These results indicated that ALV-J infection could trigger the caspase-1- mediated inflammatory response in chicks. Thus, an understanding of the inflammatory responses can provide a better insight into the pathogenicity of ALV-J and a possible anti-virus target for ALV-J infection.

Effect of human interferon-λ1 recombinant adenovirus on a gastric cancer orthotopic transplantation model.

The aim of the present study was to investigate the effect of human interferon-λ1 recombinant adenovirus (r-Ad-hIFN-λ1) on gastric carcinoma. Human SGC-7901 cells were utilized to create an orthotopic implantation model of gastric cancer in nude mice through sterile surgery. The mice were randomly divided into three groups: Phosphate-buffered saline control (blank), adenovirus encoding bacterial ß-galactosidase (Ad-Lac Z) empty vector and r-Ad-hIFN-λ1. Tumor size was measured every seven days. After three weeks of treatment, the tumors in the mice were detected by abdominal B ultrasound. The cDNA of IFN-λ1 expression in skeletal muscle was detected by a reverse transcription polymerase chain reaction and IFN-λ1 protein expression in the tumors was detected by western blot analysis and immunohistochemistry. Flow cytometry and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assays were conducted to analyze the proportion of natural killer (NK) cells in the spleen and the rate of cell apoptosis in tumor paraffin sections. Prior to sacrifice, the size of the tumors in the r-Ad-hIFN-λ1, Ad-Lac Z and blank groups was 184.29±10.84 mm3, 234.62±10.59 mm3 and 253.18±7.69 mm3, respectively (P<0.001). The lymph node metastasis in the abdominal cavity was 0% in the r-Ad-hIFN-λ1 group, 50% in the Ad-Lac Z group and 80% in the blank group (P<0.005). Furthermore, IFN-λ1 mRNA and protein were highly expressed in the r-Ad-hIFN-λ1 group, and the apoptosis rate in the r-Ad-hIFN-λ1 group was higher than that in the Ad-Lac Z and blank groups. The proportion of NK cells in the spleens of nude mice in the r-Ad-hIFN-λ1, Ad-Lac Z and blank groups was 26.53±1.54, 17.70±1.09 and 16.35±1.43%, respectively (P<0.001). The TUNEL results showed there was significantly more severe apoptosis in the r-Ad-hIFN-λ1 group than that in the two other groups. The apoptosis indices in the r-Ad-hIFN-λ1, Ad-Lac Z and blank groups were 0.772±0.075, 0.329±0.169 and 0.265±0.049, respectively. In conclusion, the r-Ad-hIFN-λ1 significantly inhibited human gastric cancer, possibly by promoting apoptosis of the tumors and stimulating immunological function.

[Comparative study on effects of different acupuncture manipulation methods at neiguan (PC 6) on hand spasm in the patient of stroke].

OBJECTIVE: To observe therapeutic effects of different needling methods at Neiguan (PC 6) on hand spasm in the patient of stroke. METHODS: One hundred and seventy-two cases were randomly divided into a simple acupuncture group (n=36), a slowly twirling needle group (n = 34), a rapidly twirling needle group (n = 30), a simple lifting-thrusting group (n = 35) and a lifting-thrusting-twirling needle group (n = 37). Acupuncture was given at Neiguan (PC 6), twice each day, 5 min each time, and in combination with rehabilitation exercise. Modified Ashworth scale and Fugl-Meyer (FMA) hand function score were used for assessment of therapeutic effects. RESULTS: The transient effective rate and the effective rate after treatment for 8 weeks for hand spasm were 66.7% and 55.6% in the simple acupuncture group, 82.4% and 85.3% in the slowly twirling needle group, 96.7% and 90.0% in the rapidly twirling needle group, 80.0% and 82.9% in the simple lifting-thrusting group and 94.6% and 91.9% in the lifting-thrusting-twirling needle group, the rapidly twirling needle group and the lifting-thrusting-twirling needle group being better than other 3 groups (P<0.01). After treatment, the grades for Ashworth scale and FMA scores in the lifting-thrusting-twirling needle group and the rapidly twirling needle group were better than those in other 3 groups (P<0.05, P<0.01), with no significant difference between the two groups (P>0.05). CONCLUSION: Acupuncture at Neiguan (PC 6) has a very definite transient effect, and the lifting-thrusting-twirling needling manipulation and the rapidly twirling needling manipulation have the best inhibitive effect on spasm.