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Polyhydroxyalkanoates (PHAs) are polymers obtained by esterification of hydroxy fatty acid monomers. Due to similar mechanical characteristics of traditional petroleum-based plastics, 100% biodegradability and biocompatibility, PHAs are considered to be one of the most potential green materials. However, the application and promotion of PHAs as a green and environmentally friendly material are difficult because of the high production costs. This article focuses on the current methods to reduce production cost of PHAs effectively, such as cell morphology regulation, metabolic pathway construction, economic carbon source utilization and open fermentation technology development. Despite most research results are still limited in laboratory, the research methods and directions provide theoretical guidance for the industrial production of economic PHAs.
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Fermentation , Industry , Petroleum , Plastics , PolyhydroxyalkanoatesABSTRACT
Objective@#To compare relevant dosimetric parameters of non-coplanar volumetric modulated arc therapy (VMAT) in treating brain tumors in conventional flattening filter (FF) or flattening filter-free (FFF) delivery mode, aiming to explore the appropriate evaluation method of accelerator for stereotactic radiosurgery (SRS).@*Methods@#Clinical data of 10 patients with single cranial tumor were retrospectively analyzed. All patients received non-coplanar VMAT at a prescription dose of 25 Gy in 1 fraction. Dosimetric parameters including conformity index (CI), heterogeneity index (HI), gradient index (GI50, GI25), gradient, volume of the brain tissue receiving larger than 10 Gy and 12 Gy(V10 and V12) and beam-on time were statistically compared between two treatment plans by paired sample t-test.@*Results@#When FFF-VMAT was compared with FF-VMAT in SRS for intracranial tumors, Paddick gradient index GI50 was 2.91±0.34 vs.3.07±0.35, 6.91±0.28 vs.7.35±0.27 for GI25, (0.57±0.07) cm vs.(0.61±0.08) cm for gradient, respectively (all P<0.05), whereas CI did not significantly differ (P>0.05). For the normal brain tissues, the average dose was (160.64±43.64) cGy vs.(174.27±53.98) cGy, (45.35±30.32)% vs.(48.37±30.88)% for V10 and (36.69±25.15)% vs.(39.48±25.37)% for V12, respectively (all P<0.05).@*Conclusions@#Non-coplanar VMAT in FFF delivery mode can improve dose distribution and normal brain tissue sparing in the treatment of intracranial single tumors. Meanwhile, supplement of GI index and gradient index can implement comprehensive evaluation of the SRS planning.
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Objective To compare relevant dosimetric parameters of non-coplanar volumetric modulated arc therapy (VMAT) in treating brain tumors in conventional flattening filter (FF) or flattening filter-free (FFF) delivery mode,aiming to explore the appropriate evaluation method of accelerator for stereotactic radiosurgery (SRS).Methods Clinical data of 10 patients with single cranial tumor were retrospectively analyzed.All patients received non-coplanar VMAT at a prescription dose of 25 Gy in 1 fraction.Dosimetric parameters including conformity index (CI),heterogeneity index (HI),gradient index (GI50,GI25),gradient,volume of the brain tissue receiving larger than 10 Gy and 12 Gy (V10 and V12) and beam-on time were statistically compared between two treatment plans by paired sample t-test.Results When FFF-VMAT was compared with FF-VMAT in SRS for intracranial tumors,Paddick gradient index GI50 was 2.91±0.34 vs.3.07±0.35,6.91±0.28 vs.7.35±0.27 for GI25,(0.57±0.07) cm vs.(0.61±0.08) cm for gradient,respectively (all P＜0.05),whereas CI did not significantly differ (P＞0.05).For the normal brain tissues,the average dose was (160.64±43.64) cGy vs.(174.27± 53.98) cGy,(45.35± 30.32) ％ vs.(48.37± 30.88) ％ for V10 and (36.69±25.15) ％ vs.(39.48±25.37) ％ for V12,respectively (all P＜0.05).Conclusions Non-coplanar VMAT in FFF delivery mode can improve dose distribution and normal brain tissue sparing in the treatment of intracranial single tumors.Meanwhile,supplement of GI index and gradient index can implement comprehensive evaluation of the SRS planning.
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Dipentodon is a monotypic genus of Dipentodontaceae and the only species, Dipentodon sinicus, is scattered in southwest China as well as adjacent Myanmar, northeast India and northern Vietnam. This species was evaluated as vulnerable in ‘China Species Red List’. Here, we assembled and characterized the complete chloroplast (cp) genome of D. sinicus using Illumina sequencing data for the first time. The complete cp genome was 158,795 bp in length, consisting of a pair of inverted repeats of 26,587 bp, a large single-copy region of 87,233 bp and a small single-copy region of 18,388 bp. The genome encoded 113 unique genes, including 79 protein-coding genes, 30 tRNA genes and four rRNA genes. Phylogenetic analysis based on 16 complete cp genome sequences indicated that D. sinicus is a member of Huerteales, consistent with its position in the latest classification of flowering plants (AGP IV).
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Objective To screen the transient and stable cell lines with high production of Nectin-like 4 (Necl-4) protein. Methods First, cDNA sequences encoding the extracellular domain of Necls were cloned into the modified vector pAPtag at the N terminus of alkaline phosphatase (AP) for fusion expression. Next, 293ET cells stably expressed Necls-AP fusion protein and secreted it into the culture medium which were detected by the AP activity assay and Western blot analysis. Then, by adding N-glycosylation processing inhibitor kifunensine into the medium, complex glycan was inhibited to generate. The residual glycan of purified protein was removed by endoglycosidase H. Finally, AP protein was removed by using human rhinovirus protease and size exclusion chromatography. The concentration of purified Necl-4 protein was monitored by measuring the absorbance at 280 nm and analyzed by SDS-PAGE. Result The transient and stable cell lines with high production of Necl-4 protein were screened by the color reaction with the AP-tag in the recombinant vector. The soluble and active form of purified Necl-4 protein was obtained after deglycosylation of native N-glycan protein with an expression level of 4 mg/L culture and purity of 95%. Conclusions By using modified AP mammalian protein expression system, we can easily screen the high productive stable cell lines by using AP activity assay. By adding mannosidase inhibitor kifunensine into the medium and cutting purified protein by using endoglycosidase H, we can obtain deglycosylated Necl-4 protein in milligram quantities. Our method might throw a light on the expression and purification of glycoprotein for structural and functional studies.
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Different types of acids from fermentation environment or industrial microorganisms exist during fermentation process. Acids may inhibit growth and metabolism of industrial strains, namely acid stress. The tolerance mechanisms of acid stress include regulation of intracellular proton concentration, protection and restoration of intracellular macromolecules, changes in cell membrane composition and acid stress response at whole cell level. Screening and modification methods have been applied to improve acid-stress tolerance of industrial strains for decades. In this review, we provide insights into acid-stress tolerance of industrial microorganisms and address the modification of microbial pathways to improve acid-stress tolerance.
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Acids , Chemistry , Fermentation , Hydrogen-Ion Concentration , Industrial Microbiology , Metabolic EngineeringABSTRACT
<p><b>OBJECTIVE</b>To investigate the molecular mechanism of nectin-like molecule 1 (NECL1) inhibiting the migration and invasion of U251 glioma cells.</p><p><b>METHODS</b>We infected U251 glioma cells with adeno-nectin-like molecule 1 (Ad-NECL1) or empty adenovirus (Ad). Transwell and wound healing assays were performed to observe the migration of U251 cells incubated with the cell supernatant from Ad-NECL1 or Ad infected U251 cells. DNA microarray was applied to screen the gene expression profile after the restoration of NECL1 in U251 glioma cell lines. The differential expression of osteopontin (OPN), a gene related to migration and invasion, was further analyzed with semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and immunohistochemistry.</p><p><b>RESULTS</b>The restoration of NECL1 inhibited migration of U251 cells significantly (P<0.05). Altogether 195 genes were found differentially expressed by microarray, in which 175 were up-regulated and 20 down-regulated, including 9 extracellular matrix proteins involved in the migration of cells. Both mRNA and protein expressions of OPN, the most markedly reduced extracellular matrix protein, were found decreased in U251 cells after restoration of NECL1. Immunohistochemical assay also detected an increase of OPN in glioma tissues, related with the progressing of malignant grade.</p><p><b>CONCLUSION</b>A link might exist between NECL1 and the extracellular matrix protein OPN in inhibiting the migration and invasion of U251 glioma cells.</p>