Modification was made on the reversed-phase high performance liquidchromatography(RP-HPLC)with Yue et al’s method as a base.The modified RP-HPLCwas used to detect leukotriene B<sub>4</sub>(LTB<sub&g...Modification was made on the reversed-phase high performance liquidchromatography(RP-HPLC)with Yue et al’s method as a base.The modified RP-HPLCwas used to detect leukotriene B<sub>4</sub>(LTB<sub>4</sub>)and 5-hydroxyeicosatetraenoic acid(5-HETE).Itwas found that the modified method has the merits of simpler procedures,shortertesting time and more satisfactory efficacy.展开更多
目的对叶酸、维生素B_(12)(VB_(12))、二甲双胍联用对2型糖尿病(T2DM)患者的治疗作用。方法选取2021年2月至2023年3月浙江省医疗健康集团长兴医院收治的112例T2DM患者,采用随机数字表法分为对照组(VB_(12)^(+)二甲双胍,56例)和研究组(叶...目的对叶酸、维生素B_(12)(VB_(12))、二甲双胍联用对2型糖尿病(T2DM)患者的治疗作用。方法选取2021年2月至2023年3月浙江省医疗健康集团长兴医院收治的112例T2DM患者,采用随机数字表法分为对照组(VB_(12)^(+)二甲双胍,56例)和研究组(叶酸^(+)VB_(12)^(+)二甲双胍,56例)。对比血糖、糖化血红蛋白(HbA1c)、空腹胰岛素(FINS)水平、胰岛素抵抗指数(HOMA-IR)和同型半胱氨酸(Hcy)水平、临床疗效、CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+)水平、Th1、Th17、Th1/Th17水平,不良反应及并发症。结果治疗后,研究组空腹血糖(FPG)、餐后2 h血糖(2 h PG)、HOMA-IR、HbA1c、FINS和Hcy均低于治疗前与对照组(P<0.05);研究组总有效率更高(P<0.05);治疗后研究组CD4^(+)、CD4^(+)/CD8^(+)均高于治疗前和对照组(P<0.05),CD8^(+)低于治疗前和对照组(P<0.05);治疗后,研究组Th1、Th17水平低于治疗前和对照组(P<0.05),Th1/Th17高于治疗前和对照组(P<0.05);2组患者不良反应发生率比较差异无统计学意义(P>0.05);研究组并发症发生率低于对照组(P<0.05)。结论叶酸与VB_(12)、二甲双胍联用治疗T2DM有助于控糖,还可提高免疫,调节Th1/Th17平衡,且安全可靠,有助于减少并发症。展开更多
△^12 fatty acid desaturase gene has been targeted as a logical candidate controlling the high oleate trait in peanut seeds. By RT-PCR method, the full-length cDNAs of △^12 fatty acid desaturase gene were isolated fr...△^12 fatty acid desaturase gene has been targeted as a logical candidate controlling the high oleate trait in peanut seeds. By RT-PCR method, the full-length cDNAs of △^12 fatty acid desaturase gene were isolated from peanut (Arachis hypogaea L.) genotypes with normal and high ratio of oleic to linoleic acid, which were designated AhFAD2B and AhFAD2B', respectively. Sequence alignment of their coding regions revealed that an extra A was inserted at the position +442 bp of AhFAD2B' sequence of high oleic acid genotypes, which resulted in the shift of open reading frame and a truncated protein AhFAD2B', with the loss of one histidine box involved in metal ion complex required for the reduction of oxygen. Analysis of transcript level showed that the expression of △^12 fatty acid desaturase gene in high oleic acid genotype was slightly lower than that in normal genotype. The enzyme activity experiment of yeast (Saccharomyces cerevisiae) cell transformed with AhFAD2B or AhFAD2B' proved that only AhFAD2B gene product showed significant △^12 fatty acid desaturase activity, but AhFAD2B' gene product did not. These results suggested that the change of AhFAD2B' gene sequence resulted in lower activity or deactivation of △^12 fatty acid desaturase in high oleic acid genotype.展开更多
A reversed phase high performance liquid chromatography (HPLC) method was established for the simultaneous determination of 12, 13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture. A Grace Apollo Cl8 col...A reversed phase high performance liquid chromatography (HPLC) method was established for the simultaneous determination of 12, 13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture. A Grace Apollo Cl8 column (250 mm × 4.6 mm, 5 μm) was used as the stationary phase and the mobile phase was composed of acetonitrile and aqueous phosphoric acid (0.2%, v/v). Gradient elution was carried out at the flow rate of 1.0 mL/min and the column temperature was 30 ℃. An ultraviolet (UV) detector was used with a selected wavelength of 240 nm. Calibration curves were linear within the concentration range of 4.6-45.75 μg/mL for 12, 13-dihydroxyeuparin (r〉0.9999) and 106.9-1068.9μg/mL for glycyrrhizic acid (r〉0.9999), respectively. Recoveries were 102.18% for 12, 13-dihydroxyeuparin and 101.17% for glycyrrhizic acid. The method developed could be applied to the simultaneous determination of 12, 13- dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.展开更多
Synthesis of acetic acid by direct oxidation of ethylene on Pd-H4SiW12O40-based catalysts was studied in a fixed-bed integral reactor and a pulse differential reactor. From the performance of the catalysts with differ...Synthesis of acetic acid by direct oxidation of ethylene on Pd-H4SiW12O40-based catalysts was studied in a fixed-bed integral reactor and a pulse differential reactor. From the performance of the catalysts with different compositions and configurations, it is proposed that acetic acid is predominantly produced via an intermediate of acetaldehyde. This can be easily confirmed by comparing the product distributions in the integral and the differential reactors. The active sites for acetic acid formation are considered to exist mainly at the boundaries between the H4SiW12O40 and the Pd particles. The Pd-based catalysts reduced by H2/N2 have higher activities than those reduced by hydrazine, as explained by the degree of Pd dispersion obtained from the characteristics of hydrogen chemical adsorption. It was found that the Pd-Se-SiW12/SiO2 catalyst with selenium tetrachloride as a precursor was more active than that with potassium selenite, and that the acetic acid yield can be greatly increased by adding a suitable amount of dichloroethane (C2H4C12/C2H4 mole ratio=0.03) to the reactants.展开更多
The cDNA of the delta-12 fatty acid desaturase gene, IgFAD2, was cloned from the marine microalgae Isochrysis galbana, a species capable of producing docosahexaenoic acid. Sequence analysis indicated that the open rea...The cDNA of the delta-12 fatty acid desaturase gene, IgFAD2, was cloned from the marine microalgae Isochrysis galbana, a species capable of producing docosahexaenoic acid. Sequence analysis indicated that the open reading frame measured a length of 1 158 bp and encoded 386 amino acids with a predicted molecular weight of 42.8 kDa and an isoelectric point of 9.2. Computational analysis of the protein sequence of IgFAD2 showed typical features of membrane-bound desaturase such as three conserved histidine boxes along with four membranespanning regions that were universally present among plant desaturases. Quantitative real-time PCR results showed that the abundance of IgFAD2 transcript was significantly upregulated under different environmental stresses including low temperature(15℃), high salinity(salinity of 62 and 93), and nitrogen starvation(220 μmol/L). Heterologous expression indicated that yeast cells transformed with a plasmid construct containing IgFAD2 could convert endogenous oleic acid(18:1^(?9), OA) into linoleic acid(18:2^(?9, 12), LA). These findings confirm that I. galbana IgFAD2 plays important roles in the biosynthetic pathways of unsaturated fatty acids.展开更多
The freshwater microalga Haematococcus pluvialis accumulates large amounts of fatty acids in response to adverse conditions.However,the key fatty acid desaturase genes in H.pluvialis remain unknown.In this study,we cl...The freshwater microalga Haematococcus pluvialis accumulates large amounts of fatty acids in response to adverse conditions.However,the key fatty acid desaturase genes in H.pluvialis remain unknown.In this study,we cloned and functionally characterized aΔ12 fatty acid desaturase gene,and designated it as HpFAD2.The open reading frame of HpFAD2 consisted of 1137 base pairs and encoded 378 amino acids.The deduced polypeptide showed 70%identity to other endoplasmic reticulumΔ12 fatty acid desaturases,whereas it had only 44%identity to plastidΔ12 fatty acid desaturases.The PSORT algorithm and phylogenetic analysis further confirmed its affiliation to the endoplasmic reticulumΔ12 fatty acid desaturases.Heterologous expression was performed in Saccharomyces cerevisiae cells transformed with the recombinant plasmid pYES2-HpFAD2.Two additional fatty acids(C16:2 and C18:2)were detected in the yeast transformants.The results indicatedΔ12 desaturation activity and substrate preference for C18:1 over C16:1.The transcriptional levels of H.pluvialis HpFAD2 at different growth stages were measured by quantitative polymerase chain reaction(PCR),indicating that the HpFAD2 transcriptional levels were significantly higher in red cells than those in green cells.Our study brings more insight into the fatty acid biosynthetic pathway of H.pluvialis.展开更多
文摘Modification was made on the reversed-phase high performance liquidchromatography(RP-HPLC)with Yue et al’s method as a base.The modified RP-HPLCwas used to detect leukotriene B<sub>4</sub>(LTB<sub>4</sub>)and 5-hydroxyeicosatetraenoic acid(5-HETE).Itwas found that the modified method has the merits of simpler procedures,shortertesting time and more satisfactory efficacy.
文摘目的对叶酸、维生素B_(12)(VB_(12))、二甲双胍联用对2型糖尿病(T2DM)患者的治疗作用。方法选取2021年2月至2023年3月浙江省医疗健康集团长兴医院收治的112例T2DM患者,采用随机数字表法分为对照组(VB_(12)^(+)二甲双胍,56例)和研究组(叶酸^(+)VB_(12)^(+)二甲双胍,56例)。对比血糖、糖化血红蛋白(HbA1c)、空腹胰岛素(FINS)水平、胰岛素抵抗指数(HOMA-IR)和同型半胱氨酸(Hcy)水平、临床疗效、CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+)水平、Th1、Th17、Th1/Th17水平,不良反应及并发症。结果治疗后,研究组空腹血糖(FPG)、餐后2 h血糖(2 h PG)、HOMA-IR、HbA1c、FINS和Hcy均低于治疗前与对照组(P<0.05);研究组总有效率更高(P<0.05);治疗后研究组CD4^(+)、CD4^(+)/CD8^(+)均高于治疗前和对照组(P<0.05),CD8^(+)低于治疗前和对照组(P<0.05);治疗后,研究组Th1、Th17水平低于治疗前和对照组(P<0.05),Th1/Th17高于治疗前和对照组(P<0.05);2组患者不良反应发生率比较差异无统计学意义(P>0.05);研究组并发症发生率低于对照组(P<0.05)。结论叶酸与VB_(12)、二甲双胍联用治疗T2DM有助于控糖,还可提高免疫,调节Th1/Th17平衡,且安全可靠,有助于减少并发症。
文摘△^12 fatty acid desaturase gene has been targeted as a logical candidate controlling the high oleate trait in peanut seeds. By RT-PCR method, the full-length cDNAs of △^12 fatty acid desaturase gene were isolated from peanut (Arachis hypogaea L.) genotypes with normal and high ratio of oleic to linoleic acid, which were designated AhFAD2B and AhFAD2B', respectively. Sequence alignment of their coding regions revealed that an extra A was inserted at the position +442 bp of AhFAD2B' sequence of high oleic acid genotypes, which resulted in the shift of open reading frame and a truncated protein AhFAD2B', with the loss of one histidine box involved in metal ion complex required for the reduction of oxygen. Analysis of transcript level showed that the expression of △^12 fatty acid desaturase gene in high oleic acid genotype was slightly lower than that in normal genotype. The enzyme activity experiment of yeast (Saccharomyces cerevisiae) cell transformed with AhFAD2B or AhFAD2B' proved that only AhFAD2B gene product showed significant △^12 fatty acid desaturase activity, but AhFAD2B' gene product did not. These results suggested that the change of AhFAD2B' gene sequence resulted in lower activity or deactivation of △^12 fatty acid desaturase in high oleic acid genotype.
基金supported by Guangdong Natural Product Reference Material Research & Development Central Lab,the First Affiliate Hospital of Sun Yat-sen University and the Industry-University-Research Cooperation Program from Science and Technology Department of Guangdong Province (No:2010B090400533)
文摘A reversed phase high performance liquid chromatography (HPLC) method was established for the simultaneous determination of 12, 13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture. A Grace Apollo Cl8 column (250 mm × 4.6 mm, 5 μm) was used as the stationary phase and the mobile phase was composed of acetonitrile and aqueous phosphoric acid (0.2%, v/v). Gradient elution was carried out at the flow rate of 1.0 mL/min and the column temperature was 30 ℃. An ultraviolet (UV) detector was used with a selected wavelength of 240 nm. Calibration curves were linear within the concentration range of 4.6-45.75 μg/mL for 12, 13-dihydroxyeuparin (r〉0.9999) and 106.9-1068.9μg/mL for glycyrrhizic acid (r〉0.9999), respectively. Recoveries were 102.18% for 12, 13-dihydroxyeuparin and 101.17% for glycyrrhizic acid. The method developed could be applied to the simultaneous determination of 12, 13- dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.
文摘Synthesis of acetic acid by direct oxidation of ethylene on Pd-H4SiW12O40-based catalysts was studied in a fixed-bed integral reactor and a pulse differential reactor. From the performance of the catalysts with different compositions and configurations, it is proposed that acetic acid is predominantly produced via an intermediate of acetaldehyde. This can be easily confirmed by comparing the product distributions in the integral and the differential reactors. The active sites for acetic acid formation are considered to exist mainly at the boundaries between the H4SiW12O40 and the Pd particles. The Pd-based catalysts reduced by H2/N2 have higher activities than those reduced by hydrazine, as explained by the degree of Pd dispersion obtained from the characteristics of hydrogen chemical adsorption. It was found that the Pd-Se-SiW12/SiO2 catalyst with selenium tetrachloride as a precursor was more active than that with potassium selenite, and that the acetic acid yield can be greatly increased by adding a suitable amount of dichloroethane (C2H4C12/C2H4 mole ratio=0.03) to the reactants.
基金The Basic Scientific Fund for National Public Research Institutes of China under contract No.2017Q09the Aoshan Science and Technology Innovation Project of Pilot National Laboratory for Marine Science and Technology(Qingdao)under contract No.2016ASKJ02+3 种基金the National Natural Science Foundation of China-Shandong Joint Funded Project under contract No.U1606404the National Natural Science Foundation of China under contract Nos 41776176 and 41806201the 973 Project from Chinese Ministry of Science and Technology under contract No.2015CB755904the Shandong Provincial Natural Science Foundation under contract No.ZR2015PD003
文摘The cDNA of the delta-12 fatty acid desaturase gene, IgFAD2, was cloned from the marine microalgae Isochrysis galbana, a species capable of producing docosahexaenoic acid. Sequence analysis indicated that the open reading frame measured a length of 1 158 bp and encoded 386 amino acids with a predicted molecular weight of 42.8 kDa and an isoelectric point of 9.2. Computational analysis of the protein sequence of IgFAD2 showed typical features of membrane-bound desaturase such as three conserved histidine boxes along with four membranespanning regions that were universally present among plant desaturases. Quantitative real-time PCR results showed that the abundance of IgFAD2 transcript was significantly upregulated under different environmental stresses including low temperature(15℃), high salinity(salinity of 62 and 93), and nitrogen starvation(220 μmol/L). Heterologous expression indicated that yeast cells transformed with a plasmid construct containing IgFAD2 could convert endogenous oleic acid(18:1^(?9), OA) into linoleic acid(18:2^(?9, 12), LA). These findings confirm that I. galbana IgFAD2 plays important roles in the biosynthetic pathways of unsaturated fatty acids.
基金This study was supported by the Zhejiang Provincial Natural Science Foundation of China(No.LQ16D060001)the National Natural Science Foundation of China(No.41606163)+3 种基金the Natural Science Foundation of the Ningbo Government(No.2017A610288)the Ningbo Science and Technology Research Projects,China(No.2019B10006)the Zhejiang Major Science Project,China(No.2019C02057)the Earmarked Fund for Modern Agro-Industry Technology Research System,China(No.CARS-49)and partly sponsored by K.C.Wong Magna Fund at Ningbo University.
文摘The freshwater microalga Haematococcus pluvialis accumulates large amounts of fatty acids in response to adverse conditions.However,the key fatty acid desaturase genes in H.pluvialis remain unknown.In this study,we cloned and functionally characterized aΔ12 fatty acid desaturase gene,and designated it as HpFAD2.The open reading frame of HpFAD2 consisted of 1137 base pairs and encoded 378 amino acids.The deduced polypeptide showed 70%identity to other endoplasmic reticulumΔ12 fatty acid desaturases,whereas it had only 44%identity to plastidΔ12 fatty acid desaturases.The PSORT algorithm and phylogenetic analysis further confirmed its affiliation to the endoplasmic reticulumΔ12 fatty acid desaturases.Heterologous expression was performed in Saccharomyces cerevisiae cells transformed with the recombinant plasmid pYES2-HpFAD2.Two additional fatty acids(C16:2 and C18:2)were detected in the yeast transformants.The results indicatedΔ12 desaturation activity and substrate preference for C18:1 over C16:1.The transcriptional levels of H.pluvialis HpFAD2 at different growth stages were measured by quantitative polymerase chain reaction(PCR),indicating that the HpFAD2 transcriptional levels were significantly higher in red cells than those in green cells.Our study brings more insight into the fatty acid biosynthetic pathway of H.pluvialis.