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Additional file 2: Table S1. The initial screening of high-yield citric acid transformants.

Additional file 1: Figure S1. The recombinant plasmid map of p20 and p21. (a) The recombinant plasmid (p20) of overexpression HGT1 transcripted by glaA promoter (PglaA); (b) The recombinant plasmid (p21) of overexpression HGT1 transcripted by aox1 promoter (Paox1). Figure S2. Verification of HGT1 gene overexpression plasmids and strains. (a) Verification of the plasmid p20, M: DNA marker III, 1: hyg (1389 bp), 2: PglaA -HGT1 (3312 bp), 3: HGT1-hyg (3217 bp), 4: PglaA–hyg (4710 bp); (b) Verification of the plasmid p21, M: DNA marker III, 1: Paox1-hyg (4612 bp), 2: HGT1-hyg (3217 bp), 3: hyg (1389 bp), 4: Paox1-HGT1 (3223 bp); (c) Verification of A. tumefaciens p20 and p21, M: DNA marker III, 1: p20 plasmid control, 2-3: A. tumefaciens p20 verifying of PglaA-HGT1 (3312 bp), 4: p21 plasmid control, 5-6: A. tumefaciens p21 verifying of Paox1-HGT1 (3217 bp); (d) Verification of A. niger p20 transformants, M: DNA marker III, 1: p20 plasmid control, 2: negative control, 3-10: p20 transformants genome verifying of PglaA-HGT1 (3312 bp); (e) Verification of A. niger p21 transformants M: DNA marker III, 1: p21 plasmid control, 2: negative control, 3-10: p21 transformants genome verifying of Paox1-HGT1(3217 bp); (f) HTG1-hyg verifation M: DNA marker III 1: plasmid control, 2: negative control, 2-10: PCR product of HTG1-hyg (3217 bp) from transformants genome; (g) Verifying from ku70 upstream to downstream of the transformants M: DNA marker III 1: plasmid control (5710 bp), 2 negative control (1800 bp), 3-10: transformants genome ku70 amplification. Figure S3. Multiple sequence alignment of glucose transporters. Figure S4. Validating hyg gene in the HGT1 overexpression mutants at the 15th generation. M: DNA marker III, 1: positive control (1389 bp), 2: negative control, 3-10: amplified hyg gene using the genomic DNA of overexpressed mutants. Genomes of all strains were gained from a 15-generation subculture. Figure S5. Measurement of diameter size of mycelial balls of citric acid in the 30 L bioreactor fermentation.

Hepatocellular carcinoma (HCC) is one of the most prevalent and lethal cancers. It has been demonstrated that aberrant expression of miRNAs plays an important role in HCC development. Here, we observed decreased miR-125b expression status in tumor samples from HCC patients, and the five years survival rate of HCC patients with low miR-125b expression is poor. By using bioinformatics prediction tools combining with luciferase reporter assay, we identified that miR-125b can suppress the expression of SIRT6 by directly targeting the seed-matching region of its 3'UTR. Based on the analysis via TCGA and clinical samples data, the expression of SIRT6 showed negatively correlated with the expression of mir-125b. After knocking-out the expression of SIRT6 through CRISPR/Cas9, HCC cells showed the decreased cell viability and invasiveness, which had the similar function upon the overexpression of the miR-125b. The function induced by overexpression of miR-125b can be rescued by the restoration of SIRT6. Further experiments demonstrated that the HCC cells showed the significant cellular senescence and apoptosis upon overexpression of miR-125b or knockout SIRT6, which is in accordance with the compromised cell malignancy. Thus, we conclude that, by targeting SIRT6, miR-125b can function as a tumor suppressor to induce the cellular senescence and apoptosis in hepatocellular carcinogenesis and could provide a novel insight for HCC treatment.

Aiming at the implement of RSA algorithm, the attack methods are variety. In order to ensure the algorithm can against the side channel attack, in this paper, we present a masking scheme for RSA decryption. Our scheme exploits the multiply-homomorphic property of the existing RSA encryption scheme to compute an multiply-mask as an encryption of a random message and randomly splits the secret key into two shares as the sub-calculate such that each share is statistically independent from the original value. Our solution differs in several aspects from the recent masking RSA implementation. According to encrypted random number, the original ciphertext is blinded and the splitting secret key can reduce the size of the key and speed up the calculation of the algorithm. During the decryption, all the operations are under the masking state, therefore, through multiply masking and secret key splitting, we can secure a RSA implementation, the scheme we proposed can against the timing attack, simple power attack and differential power attack. Compared with others, this scheme can reach a higher calculation and security level.

By adding appropriate proportions of nitrobenzene (C6H5NO2) to the blended solution of poly{[4,8-bis-(2-ethyl-hexyl-thiophene-5-yl)-benzo[1,2-b:4,5-b′]dithiophene-2,6-diyl]-alt-[2-(2′-ethyl-hexanoyl)-thieno[3,4-b]thiophen-4,6-diyl]}:[6,6]-phenyl-C71-butyric acid methyl ester (PBDTTT-C-T: PC71BM), we substantially improved the power conversion efficiency from the best reported value of 7.48–8.88%. Experiments and simulations show that nitrobenzene and PBDTTT-C-T form stable coplanar charge-transfer complexes through hydrogen bonds. Formation of the PBDTTT-C-T-C6H5NO2 complex simultaneously increases the external quantum efficiency. The underlying mechanisms of increased external quantum efficiency are attributed to the following: (i) higher lowest unoccupied molecular orbital (LUMO) of PBDTTT-C-T-C6H5NO2 for more efficient photoinduced electron transfer to the LUMO of PC71BM and (ii) efficient quenching of fluorescence in the active layers due to formation of the PBDTTT-C-T-C6H5NO2 complex. This discovery clearly illustrates the potential of hydrogen-bonded complexes as a new route for efficient polymer-based photovoltaic devices. Copyright © 2014 John Wiley & Sons, Ltd.

The high potential for occurrence and the negative consequences of secondary accidents make them an issue of great concern affecting freeway safety. Using accident records from a three-year period together with California interstate freeway loop data, a dynamic method for more accurate classification based on the traffic shock wave detecting method was used to identify secondary accidents. Spatio-temporal gaps between the primary and secondary accident were proven be fit via a mixture of Weibull and normal distribution. A logistic regression model was developed to investigate major factors contributing to secondary accident occurrence. Traffic shock wave speed and volume at the occurrence of a primary accident were explicitly considered in the model, as a secondary accident is defined as an accident that occurs within the spatio-temporal impact scope of the primary accident. Results show that the shock waves originating in the wake of a primary accident have a more significant impact on the likelihood of a secondary accident occurrence than the effects of traffic volume. Primary accidents with long durations can significantly increase the possibility of secondary accidents. Unsafe speed and weather are other factors contributing to secondary crash occurrence. It is strongly suggested that when police or rescue personnel arrive at the scene of an accident, they should not suddenly block, decrease, or unblock the traffic flow, but instead endeavor to control traffic in a smooth and controlled manner. Also it is important to reduce accident processing time to reduce the risk of secondary accident.

Pterostilbene (trans-3,5-dimethoxy-4ʹ-hydroxystilbene) is an important phenolic compound which is reported as a potential anti-oxidative, anti-inflammatory, and anti-cancer agent. It is a natural compound and belongs to the group of phytoalexins, agents produced by plants to fight infections like chemicals and fungal. In nature, pterostilbene can be oxidized by peroxidase from fungus, and peroxidase also exists in human body. So it may undergo oxidative process in human as well. 3ʹ-hydroxypterostilbene is another natural phenolic compound and also is a metabolite of pterostilbene identified in mouse. The two adjacent hydroxyl groups in its molecule indicate that it may have a stronger bioactivity than pterostilbene. To understand the possible oxidative reaction and major products in metabolic process, this research use peroxidase and phosphate-citrate buffer to simulate the peroxidation of pterostilbene and 3ʹ-hydroxypterostilbene in vitro. The major oxidative products were identified as pterostilbene trans dehydrodimer and 3ʹ-hydroxypterostilbene trans dehydrodimer by LC/MS and NMR. The anti-inflammatory and anti-cancer cell proliferative activity of three compounds, pterostilbene, 3ʹ-hydroxypterostilbene, and pterostilbene trans dehydrodimer, was evaluated in this research. In anti-inflammatory evaluation, Griess assay was used to measure the nitric oxide production in RAW264.& cells which were treated with the three compounds at different concentration. In anti-cancer cell proliferative study, MTT assay was used to measure the cell viability of two cancer cell lines: drug-sensitive HL-60 leukemia cell line and drug-resistant MDA-MB231 breast cancer cell line. According to our data, 3ʹ-hydroxypterostilbene had a significant anti-inflammatory and anti-cancer cell proliferative activity.