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Additional file 1: Table S1. Sequences of primers used in this study. Table S2. Medium composition. Table S3. Comparison of codon usage. Figure S1. Structure of the vector p35S-ZmNrt used for tobacco transformation. Figure S2. Comparison of the amino acid sequences of the maize high-affinity nitrate transporter ZmNrt2.1 and the tobacco high-affinity nitrate transporter NtNrt2.1. Figure S3. PCR analysis of genomic DNA extracted from leaves of wild type and putative transgenic tobacco plants. Figure S4. RT-PCR screening for transgenic plants. Figure S5. Gene expression analysis. Figure S6. Soluble nitrate contents in plant shoots and roots. Figure S7. Representative photographs of plant growth with various levels of nitrate, Ca2+ and K+. Figure S8. NtNAR gene expression analysis.

Additional file 1: Figure S1. Amino acid sequences multiple alignment of 2-HADH from P. aeruginosa CCTCC M 2011394, B. xenovorans LB400, P. putida ATCC 12633, P. aeruginosa NUST, P. fluorescens strain EBC191. Figure S2. Optimization of biooxidation reaction conditions by resting cells of recombinant E. coli BL21(DE3)/pET28b-HADH. (A) Effect of temperature on the biooxidation. The optimum temperature was determined over the range from 25 and 65 °C. The recombinant E. coli BL21(DE3)/pET28b-HADH showed high activity at 35–55 °C, and at higher temperatures the activity began to decrease significantly. (B) Effect of pH on the biooxidation; The optimum pH on the oxidation reaction was determined over the range from 6.0 and 9.0. When the pH was below 7.5 or over 8.5, the enzyme activity decreased dramatically. Figure S3. SDS-PAGE analysis of the expressed (R)-2-KAR in recombinant E. coli BL21(DE3)/pET28b-KAR. 1. Markers; 2. Cell-free extract of recombinant E. coli BL21(DE3)/pET28b-KAR; 3. Purified (R)-2-KAR from E. coli BL21(DE3)/pET28b-KAR (~32 kDa). Figure S4. SDS-PAGE analysis of the expressed GDH in recombinant E. coli BL21(DE3)/pET28b-GDH and coexpressed (R)-2-KAR and GDH in recombinant E. coli BL21(DE3)/pCDFDuet-KAR-GDH. 1. E. coli BL21(DE3)/pCDFDuet-KAR-GDH with 0.1 mM IPTG. The upper arrow indicated (R)-2-KAR (~32 kDa) and the lower arrow represented GDH (~28 kDa).2. E. coli BL21(DE3)/pCDFDuet-KAR-GDH with 0 mM IPTG; 3. E. coli BL21(DE3)/pET28b-GDH with 0.1 mM IPTG; 4. E. coli BL21(DE3)/pET28b-GDH with 0 mM IPTG; 5.Empty plasmid of pET28b with 0 mM IPTG. 6. Markers. Figure S5. SDS-PAGE analysis of the coexpressed (S)-2-HADH, (R)-2-KAR and GDH in recombinant E. coli BL21(DE3)/pET28b-HADH/pCDFDuet-KAR-GDH. 1. Markers; 2. Cell-free extract of recombinant E. coli BL21(DE3)/pET28b-HADH/pCDFDuet-KAR-GDH. The upper arrow indicated (S)-2-HADH (~42 kDa), the middle arrow indicated (R)-2-KAR (~32 kDa) and the lower arrow represented GDH (~28 kDa).

The aim of this study was to evaluate the short- and long-term outcomes after total gastrectomy (TG) with D2 lymphadenectomy. Patients undergoing TG with D2 lymphadenectomy for gastric cancer between December 2008 and December 2011 were enrolled. Univariate and multivariate analyses were performed to evaluate the risk factors for the short- and long-term outcomes. A total of 229 patients were analyzed, and 22.3 % developed complications within 30 days of surgery. No patient died within 30 days, while 2.6 % died within 90 days of the operation. In the multivariate analysis, age ≥65 years and cardiopulmonary comorbidities were associated with morbidity, whereas hypoproteinemia and tumor–node–metastasis (TNM) stage III were associated with the disease-free survival (DFS) and overall survival (OS). The number of preoperative risk factors stratified the morbidity from 10.3 % in those without any risk factors to 40.5 % in patients with both risk factors. Similarly, 5-year survival rates decreased from 68.9 % (DFS) and 71.1 % (OS) in those without risk factors to 20.2 % (DFS) and 22.9 % (OS) in patients with both risk factors. TG with D2 lymphadenectomy has acceptable short- and long-term outcomes. Patient risk stratification may allow for more rational selection of patients and therapeutic strategies for gastric resection.

The degree of postoperative ileus and the underlying pathophysiological mechanism among different types of surgical traumas have not been examined. The aim of this study was to investigate the inflammatory and oxidative stress changes in rat intestinal muscularis and gastrointestinal transit among three types of surgical traumas. Rats were randomized assigned to four groups: control group, intestinal manipulation (IM) group, intestinal ischemia/reperfusion injury (IR) group and peritoneal air exposure (AE) group. Gastrointestinal transit was measured 24 hours after surgery. Malondialdehyde (MDA), glutathione (GSH) and inflammatory mediators in intestinal muscularis were measured. Influx of neutrophil in intestinal muscularis was also determined. The degree of gastrointestinal motility impairment was equal between the IM and AE groups. However, the IR group was subject to a less impairment of gastrointestinal motility compared with the IM and AE groups. The IM group showed the most significant increase of inflammatory response, while the AE group showed the most significant increase of oxidative stress. The IR group showed a moderate increase of inflammatory response and oxidative stress. Rats subjected to IM, IR and AE could all develop into POI. We speculate that oxidative stress should be an equally important pathophysiological mechanism of POI as inflammation.

Aim Recent studies have shown that sarcopenia is associated with negative postoperative outcomes. However, none of these studies analysed muscle strength or physical performance, which are also important components of sarcopenia. The present study aimed to investigate whether sarcopenia itself, as defined by low muscle mass, strength and physical performance, would predict complications after surgery for colorectal cancer. Method We conducted a prospective study of patients who underwent surgery for colorectal cancer at our department between August 2014 and February 2015. Sarcopenia was diagnosed by a combination of third lumbar vertebra muscle index (L3 MI), handgrip strength and 6-m usual gait speed. Univariate and multivariate analyses evaluating the risk factors for postoperative complications were performed. Only complications classified as Grade II or above according to the Clavien–Dindo classification were analysed in this study. Results A total of 142 patients were included in the study, and 17 patients were diagnosed as having sarcopenia. Postoperative complications of Grade II or above occurred in 40 patients, including 10 with sarcopenia and 30 without sarcopenia. Multivariate analysis showed that sarcopenia and previous abdominal surgery were independent risk factors for postoperative complications. Patients with sarcopenia also had an obvious tendency to a higher incidence of infectious complications. By comparing two logistic regression models, sarcopenia showed a better predictive power for postoperative complications than did low muscle mass. Conclusion Sarcopenia and previous abdominal surgery are independent risk factors for complications after surgery for colorectal cancer. Including a functional aspect to the definition of sarcopenia may result in a better prediction of postoperative complications.

Unlike the conventional control of AC and DC motors, an SR motor requires particular control strategies to adjust its conduction angle variables for optimum operation. A fixed conduction angle control cannot match these control requirements. In this paper, a new control strategy which can move the conduction angles arbitrarily for different operation requirements is proposed. Some experimental results show that the control strategy has such functions that it can alter the excitation modes and select the optimum conduction angles for specified operation.

A new type of linear oscillatory actuator has been developed as a drive source for liquid hydrogen pump. The distinguishing feature of the linear oscillatory actuator is that not only its structure is simple with a few parts, but also it protects the cylinder from external heat as it is mounted in the liquid hydrogen tank. In this paper, the structure and operation of the linear oscillatory actuator is described first, and then the static thrust is discussed by means of numerical analysis and experiment, both verifying a fact that the static thrust achieves desired requirements for liquid hydrogen pump.