Objective To explore the neuroprotective effects of the Shaoyao Gancao decoction(SGD)against excitatory damage in PC12 cells and the role of the Src-NR2-nNOS pathway mediation by SGD in regulatingγ-aminobutyric acid(...Objective To explore the neuroprotective effects of the Shaoyao Gancao decoction(SGD)against excitatory damage in PC12 cells and the role of the Src-NR2-nNOS pathway mediation by SGD in regulatingγ-aminobutyric acid(GABA)-glutamate(Glu)homeostasis.Methods N-Methyl-d-aspartic acid(NMDA)was used to establish a PC12 cell excitability injury model.To investigate the neuroprotective effect of SGD,a cell counting kit-8(CCK-8)assay was used to determine PC12 cell viability,Annexin V/Propidium Iodide(Annexin V/PI)double staining was used to determine PC12 cell apoptosis,and Ca^(2+)concentration was observed using laser confocal microscopy.GABA receptor agonists and antagonists were used to analyze the neuroprotective interactions betweenγ-aminobutyric acid(GABA)and NMDA receptors.Additionally,molecular biology techniques were used to determine mRNA and protein expression in the Src-NR2-nNOS pathway.We analyzed the correlations between the regulatory sites of GABA and NMDA interactions,excitatory neurotoxicity,and brain damage at the molecular level.Results NMDA excitotoxic injury manifested as a significant decrease in cell activity,increased apoptosis and caspase-3 protein expression,and a significant increase in intracellular Ca^(2+)concentration.Administration of SGD,a GABAA receptor agonist(muscimol),or a GABAB receptor agonist(baclofen)decreased intracellular Ca^(2+)concentrations,attenuated apoptosis,and reversed NMDA-induced upregulation of caspase-3,Src,NMDAR2A,NMDAR2B,and nNOS.Unexpectedly,a GABA_(A)receptor antagonist(bicuculline)and a GABA_(B)receptor antagonist(saclofen)failed to significantly increase excitatory neurotoxicity.Conclusions Taken together,these results not only provide an experimental basis for SGD administration in the clinical treatment of central nervous system injury diseases,but also suggest that the Src-NR2A-nNOS pathway may be a valuable target in excitotoxicity treatment.展开更多
Background: Methylmalonic aciduria (MMA) is a genetic disorder of aminoacid metabolism, due to mutations in methylmalonyl-CoA mutase, which leads to the accumulation of methylmalonic acid in body fluids. Patients typi...Background: Methylmalonic aciduria (MMA) is a genetic disorder of aminoacid metabolism, due to mutations in methylmalonyl-CoA mutase, which leads to the accumulation of methylmalonic acid in body fluids. Patients typically present at the age of 1 month to 1 year with dehydration, renal impairment as well as neurologic manifestations viz. seizure, encephalopathy, strokes and disease in the globus pallidi. The case: a 26-year-old man presented with severe acute on top of chronic renal disease with serum creatinine at 590 umol/L and bilateral 8 cm kidneys with thin and echogenic cortex. He had: (a) hypernatremic dehydration, metabolic acidosis and high ammonia level with (b) a history of multiple similar attacks since the age of 8 months. Diagnosis of MMA was confirmed by high serum and urine enzymatic levels as well as genetic testing. His initial management included support with replacements of fluids, electrolytes, and bicarbonates as well as intravenous dextrose, vitamin B12 and broad-spectrum antibiotic (Meropenem) for his chest infection. Subsequently, he received 1) CARBAGLU (carglumic acid) for 7 days to lower his ammonia level to Conclusion: Untreated homozygous MMA variants, can achieve adulthood with significant renal disease yet their morbidity and mortality can be ameliorated with diet and specific therapy.展开更多
Modification was made on the reversed-phase high performance liquid chromatography(RP-HPLC)with Yue et al's method as a base.The modified RP-HPLC was used to detect leukotriene B_4(LTB_4)and 5-hydroxyeicosatetraen...Modification was made on the reversed-phase high performance liquid chromatography(RP-HPLC)with Yue et al's method as a base.The modified RP-HPLC was used to detect leukotriene B_4(LTB_4)and 5-hydroxyeicosatetraenoic acid(5-HETE).It was found that the modified method has the merits of simpler procedures,shorter testing 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
A full-length sequence coding for △^12 fatty acid desaturase gene from peanut（Arachis hypogaea L.）was cloned into the expression vector, pRSETB, to generate recombinant plasmid pRSET/HO-A, which was subsequently tr...A full-length sequence coding for △^12 fatty acid desaturase gene from peanut（Arachis hypogaea L.）was cloned into the expression vector, pRSETB, to generate recombinant plasmid pRSET/HO-A, which was subsequently transformed into expression Escherichia. coli BL21（DE3）pLysS. The △^12 fatty acid desaturase was highly expressed in E. coli BL21（DE3）pLysS in the presence of isopropyl-D-thiogalactopyranoside （IPTG）. The fusion protein was purified and used to form a reaction system in vitro by adding oleic acid as substrate and incubating it at 20℃ for 6 h. Total fatty acids was extracted and methlesterified and then analyzed with gas chromatography. A novel peak corresponding to linoleic acid methyl ester standards was detected with the same retention time. GC-MS （gas chromatogram and gas chromatogram-mass spectrometry） analysis showed that the novel peak was linoleic acid methyl ester. These results exhibited △^12 fatty acid desaturase activity, which could convert oleic acid to linoleic acid specifically.展开更多
基金supported by the National Natural Science Foundation of China(82074036).
文摘Objective To explore the neuroprotective effects of the Shaoyao Gancao decoction(SGD)against excitatory damage in PC12 cells and the role of the Src-NR2-nNOS pathway mediation by SGD in regulatingγ-aminobutyric acid(GABA)-glutamate(Glu)homeostasis.Methods N-Methyl-d-aspartic acid(NMDA)was used to establish a PC12 cell excitability injury model.To investigate the neuroprotective effect of SGD,a cell counting kit-8(CCK-8)assay was used to determine PC12 cell viability,Annexin V/Propidium Iodide(Annexin V/PI)double staining was used to determine PC12 cell apoptosis,and Ca^(2+)concentration was observed using laser confocal microscopy.GABA receptor agonists and antagonists were used to analyze the neuroprotective interactions betweenγ-aminobutyric acid(GABA)and NMDA receptors.Additionally,molecular biology techniques were used to determine mRNA and protein expression in the Src-NR2-nNOS pathway.We analyzed the correlations between the regulatory sites of GABA and NMDA interactions,excitatory neurotoxicity,and brain damage at the molecular level.Results NMDA excitotoxic injury manifested as a significant decrease in cell activity,increased apoptosis and caspase-3 protein expression,and a significant increase in intracellular Ca^(2+)concentration.Administration of SGD,a GABAA receptor agonist(muscimol),or a GABAB receptor agonist(baclofen)decreased intracellular Ca^(2+)concentrations,attenuated apoptosis,and reversed NMDA-induced upregulation of caspase-3,Src,NMDAR2A,NMDAR2B,and nNOS.Unexpectedly,a GABA_(A)receptor antagonist(bicuculline)and a GABA_(B)receptor antagonist(saclofen)failed to significantly increase excitatory neurotoxicity.Conclusions Taken together,these results not only provide an experimental basis for SGD administration in the clinical treatment of central nervous system injury diseases,but also suggest that the Src-NR2A-nNOS pathway may be a valuable target in excitotoxicity treatment.
文摘Background: Methylmalonic aciduria (MMA) is a genetic disorder of aminoacid metabolism, due to mutations in methylmalonyl-CoA mutase, which leads to the accumulation of methylmalonic acid in body fluids. Patients typically present at the age of 1 month to 1 year with dehydration, renal impairment as well as neurologic manifestations viz. seizure, encephalopathy, strokes and disease in the globus pallidi. The case: a 26-year-old man presented with severe acute on top of chronic renal disease with serum creatinine at 590 umol/L and bilateral 8 cm kidneys with thin and echogenic cortex. He had: (a) hypernatremic dehydration, metabolic acidosis and high ammonia level with (b) a history of multiple similar attacks since the age of 8 months. Diagnosis of MMA was confirmed by high serum and urine enzymatic levels as well as genetic testing. His initial management included support with replacements of fluids, electrolytes, and bicarbonates as well as intravenous dextrose, vitamin B12 and broad-spectrum antibiotic (Meropenem) for his chest infection. Subsequently, he received 1) CARBAGLU (carglumic acid) for 7 days to lower his ammonia level to Conclusion: Untreated homozygous MMA variants, can achieve adulthood with significant renal disease yet their morbidity and mortality can be ameliorated with diet and specific therapy.
文摘Modification was made on the reversed-phase high performance liquid chromatography(RP-HPLC)with Yue et al's method as a base.The modified RP-HPLC was used to detect leukotriene B_4(LTB_4)and 5-hydroxyeicosatetraenoic acid(5-HETE).It was found that the modified method has the merits of simpler procedures,shorter testing 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。
基金This work was supported by Chinese National Programs for High Technology Research and Development (No. 2002AA207004).
文摘A full-length sequence coding for △^12 fatty acid desaturase gene from peanut（Arachis hypogaea L.）was cloned into the expression vector, pRSETB, to generate recombinant plasmid pRSET/HO-A, which was subsequently transformed into expression Escherichia. coli BL21（DE3）pLysS. The △^12 fatty acid desaturase was highly expressed in E. coli BL21（DE3）pLysS in the presence of isopropyl-D-thiogalactopyranoside （IPTG）. The fusion protein was purified and used to form a reaction system in vitro by adding oleic acid as substrate and incubating it at 20℃ for 6 h. Total fatty acids was extracted and methlesterified and then analyzed with gas chromatography. A novel peak corresponding to linoleic acid methyl ester standards was detected with the same retention time. GC-MS （gas chromatogram and gas chromatogram-mass spectrometry） analysis showed that the novel peak was linoleic acid methyl ester. These results exhibited △^12 fatty acid desaturase activity, which could convert oleic acid to linoleic acid specifically.
