Objective To study long-term outcome and prognostic factors of nasopharyngeal carcinoma treated by intensity modulated radiotherapy.Methods A total of 299 patients with nondisseminated nasopharyngeal carcinoma who received initial radiotherapy were analyzed retrospectively.The primary lesion and the upper neck received 70 Gy (5 fraction per week in all 30 fraction) by intensitymodulated radiotherapy (IMRT).The lower neck and the supraclavicular fossa was given 54 Gy (5 fraction per week in all 30 fraction) by a single anterior tangent field with spinal cord block.A median dose of 9.2 Gy (4-20.Gy) was given to the residual primary lesion by IMRT or X-knife.The Kaplan-Meier method was used for calculating the overall survival (OS),disease progression-free survival (DPFS),distant metastasisfree survival (DMFS),Log-rank test was used for evaluating the differences between groups.Multivariate prognostic factor was analyzcd by Cox method.Results The follow-up rate was 99.7％.119 patients were followed-up more than with 5 years.The 5-year OS for stage Ⅰ + Ⅱ,stage Ⅲ and stage Ⅳ were 97.1％,82.7％ and 52.2％(x2=46.19,P=0.000),the 5 years DPFS were 100％,77.6％ and 57.7％ (x2=23.29,P =0.000),DMFS were 100％,82.3％,63.7％ (x2 =16.57,P =0.000) respectively.The 5 year OS,DPFS and DMFS of male and female were 70.7％ vs 94.1％ (x2=16.82,P=0.000),71.5％ vs 87.3％ (x2 =4.74,P =0.029) and 77.2％ vs 89.7％ (x2 =4.38,P =0.036) respectively.For patients who were younger than 45-years,the male had a significantly unfavorable 5-year OS (66.8％ vs.91.2％,x2=7.07,P=0.008),DPFS (59.9％ vs.91.2％,x2=7.72,P=0.005) and DMFS (66.4％ vs.94.0％,x2 =8.46,P =0.004) ;For patients who were old than 45-years,only OS was significantly different between male and female (72.2％ vs.96.0％,x2 =10.19,P =0.001).Multivariate analysis showed the independent prognosticfactors for OS,DPFS,DMFS,were gender (x2 =14.27,5.72,17.64,P =0.000,0.017,0.000),TNM stage (x2 =5.33,15.70,10.57,P =0.021,0.000,0.001) and lymph nodes capsular invasion (x2 =4.30,11.08,21.24,P =0.038,0.001,0.000).Intracranial invasion and supraclavicular lymph node metastasis were independent prognostic factors for OS (x2 =13.32,5.38,P =O.000,0.020).Conclusions The TNM stage,lymph nodes capsular invasion and gender are independent prognostic factors for nasopharyngeal carcinoma treated by intensity-modulated radiotherapy.The patients of younger than 45years own a worse outcome.

To develop the stable transformants of the silkworm (Bombyx mori) BmN cells that could continuously express the exogenous gene based on a non-transposon vector, an expression cassette containing human granucyto-macrophage colony-stimulating factor (hGM-CSF) gene driven by ie-1 promoter from B. mori nucleopolyhedrovirus was inserted into pIZT-V5-His to form a recombinant vector pIZT-IE-hGM-CSF, followed by transfecting the constructant into BmN cells, the stable ie-hGM-CSF cell lines were obtained after being selected with Zeocin. PCR result using the genomic DNA of the transformed BmN cells as template illustrated a specific fragment of ie-hGM-CSF, and Western blotting analysis using an antibody against hGM-CSF demonstrated a specific band with a molecular weight of 22 kDa in the transformed cells, meanwhile, the expression level of hGM-CSF determined by ELISA was about 2 814.7 pg in 10(6) transformed BmN cells.
Animals ; Animals, Genetically Modified ; Bombyx ; cytology ; genetics ; metabolism ; Cell Line ; Fibroins ; genetics ; Genetic Vectors ; genetics ; Granulocyte-Macrophage Colony-Stimulating Factor ; biosynthesis ; genetics ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transformation, Genetic

Hydrogen production from lignocellulosic biomass is both sustainable and environmentally friendly, which is garnering more and more attention across the world, with an expectation to challenge the shortage of fossil fuels supply and climate change as well. In this article, the update research progress and technology development of biohydrogen production are reviewed, with a focus on biomass pretreatment, hydrogen-producing microorganisms and process engineering strategies. And in the meantime, a roadmap for more efficient and economic biohydrogen production is envisioned.
Bacteria ; metabolism ; Bioelectric Energy Sources ; microbiology ; trends ; Biomass ; Biotransformation ; Fermentation ; Hydrogen ; metabolism ; Lignin ; metabolism

Based on the character of strong promoter of the fibroin gene and high level secretion of fibroin of Bombyx mori, we amplified the promoter of heavy chain gene (Fib-H) and its downstream signal peptide sequence (FibHS) by PCR. After that, we cloned the PCR product in pBluescriptII SK (+) to form the vector pSK-FibHS and analyzed its sequence. The sequence identity was 99% comparable to that of the reported sequence by Blast on line. Then we digested pSk-Ser-DsRed-PolyA with Sal IKpn I to get DsRed-PolyA DNA fragment and subcloned it into vector pSK-FibHS to generate a transitorily secretory expression vector pSK-FibHS-DsRed-PolyA. After identified the recombinant plasmid by restriction enzyme digestion, we transfected pSK-FibHS-DsRed-PolyA into BmN cells by liposome. From the cells transfected with the recombinant vector, what the red fluorescence could be detected verified that the recombinant vector could express DsRed in BmN cells transiently. Furthermore, when silkworm had been injected with the recombinant vector pSK-FibHS-DsRed-PolyA, red fluorescence could be observed in the lumen of silk gland of silkworm. The result indicated that DsRed expressed transiently and was secreted into lumen of the silk gland. Therefore, we supposed that the cloned sequence (FibHS) possessed signal peptide bio-function. Moreover, this study would lay a foundation for the research on secretory expression of exogenous gene by silk gland bioreactor.
Amino Acid Sequence ; Animals ; Base Sequence ; Bombyx ; genetics ; metabolism ; Cloning, Molecular ; Fibroins ; biosynthesis ; genetics ; Insect Proteins ; biosynthesis ; genetics ; Luminescent Proteins ; biosynthesis ; genetics ; Molecular Sequence Data ; Promoter Regions, Genetic ; genetics

Objective:To evaluate the safety and efficacy of a novel domestic extracorporeal membrane oxygenation (ECMO) mainframe in a porcine model, and to provide the basis for further clinical application.Methods:Five domestic healthy male white pigs, weighing (51±4) kg, were selected. The ECMO system was established by using a novel ECMO mainframe with imported membrane oxygenator and pipeline, and continued to run for 72 hours. ECMO parameters are as follows: veno-arterial ECMO, centrifugal pump speed 3 000-3 500 r/min, continuous infusion of heparin anticoagulation to maintain the activate clotting time (ACT) of 140-200 s. Real-time monitoring of speed, flow, pressure before pump, pressure after pump, pressure after membrane and other equipment parameters, and the equipment performance was scored. The changes of hemodynamics, blood lactic acid and blood routine were monitored dynamically. Repeated measures analysis of variance was used to compare different time points within the group. At the end of the experiment, the thrombosis in the pump head and oxygenator was observed. The animals were sacrificed to obtain the tissue samples of the main organs for gross observation and pathological injury evaluation.Results:All animals successfully ran the ECMO system for 72 hours. (1) The centrifugal pump speed should be maintained at 3 029-3 483 r/min, the flow rate was maintained at 2.24-2.60 L/min, The pressure before the pump between minus 107.57 and minus 31.86 mmHg, the pressure after the pump was 197.50-282.43 mmHg, and the pressure after the membrane was 178.71-261.5 mmHg, all were in the normal range, and there was no significant difference between different time points (all P>0.05). The performance scores of the mainframe were all 4 points or above, indicating that the use requirements were met. (2) The heart rate of the animals was 50-80 beats /min, the mean arterial pressure was 85-115 mmHg, and the lactic acid was 0.996-2.25 mmol/L, all within the normal range, and there was no significant difference between different time points (all P>0.05). The free hemoglobin was 8.98-16.39 mg/L, and the hemoglobin was 6.58-7.52 g/L, both within a reasonable range, and there was no significant difference between different time points (all P>0.05). The platelet count was 69.6-231.6×10 9/L, and showed a continuous downward trend ( P<0.05). ACT was maintained at 135-169 s, which was within the target range, and there was no significant difference between different time points ( P<0.05). (3) At the end of the experiment, there was no obvious thrombosis in the pump head and oxygenator, no obvious thrombosis or infarction in the heart, brain, liver, lung and kidney, and no obvious hemorrhage or necrosis under the microscope. Conclusions:The ECMO established by the novel domestic ECMO mainframe combined with imported membrane oxygenator and pipeline ran smoothly for 72 hours, achieving the target of effect and safety.