Flavonoids have been reported to have therapeutic potential for spinal cord injury.Hawthorn leaves have abundant content and species of total flavonoids,and studies of the effects of the total flavonoids of hawthorn l...Flavonoids have been reported to have therapeutic potential for spinal cord injury.Hawthorn leaves have abundant content and species of total flavonoids,and studies of the effects of the total flavonoids of hawthorn leaves on spinal cord injury have not been published in or outside China.Therefore,Sprague-Dawley rats were used to establish a spinal cord injury model by Allen's method.Rats were intraperitoneally injected with 0.2 m L of different concentrations of total flavonoids of hawthorn leaves(5,10,and 20 mg/kg)after spinal cord injury.Injections were administered once every 6 hours,three times a day,for 14 days.After treatment with various concentrations of total flavonoids of hawthorn leaves,the Basso,Beattie,and Bresnahan scores and histological staining indicated decreases in the lesion cavity and number of apoptotic cells of the injured spinal cord tissue;the morphological arrangement of the myelin sheath and nerve cells tended to be regular;and the Nissl bodies in neurons increased.The Basso,Beattie,and Bresnahan scores of treated spinal cord injury rats were increased.Western blot assays showed that the expression levels of pro-apoptotic Bax and cleaved caspase-3 were decreased,but the expression level of the anti-apoptotic Bcl-2 protein was increased.The improvement of the above physiological indicators showed a dose-dependent relationship with the concentration of total flavonoids of hawthorn leaves.The above findings confirm that total flavonoids of hawthorn leaves can reduce apoptosis and exert neuroprotective effects to promote the recovery of the motor function of rats with spinal cord injury.This study was approved by the Ethics Committee of the Guangxi Medical University of China(approval No.201810042)in October 2018.展开更多
The structural characteristics of protein body accumulation in different endosperm regions of hard wheat cultivar （XM33） and soft wheat cultivar （NM13） under drought stress were investigated. Drought stress treatm...The structural characteristics of protein body accumulation in different endosperm regions of hard wheat cultivar （XM33） and soft wheat cultivar （NM13） under drought stress were investigated. Drought stress treatment was implemented from plant regreening to the caryopsis mature stage. Microscope images of endosperm cells were obtained using resin semi- thin slice technology to observe the distribution and relative area of protein body （PB）. Compared with NM13, relative PB area of XM33 was significantly higher in sub-aleurone endosperm region. The amount of accumulation, including the size and relative area of PB, in two wheat cultivars was higher in sub-aleurone region than that in central region at 18 days post anthesis （DPA）. Drought stress significantly enhanced the sizes and relative areas of PBs in the dorsal and abdominal endosperms in two wheat cultivars. Particularly for dorsal endosperm, drought stress enhanced the relative PB area at 18 DPA and NM13 （5.0% vs. 6.73%） showed less enhancement than XM33 （5.49% vs. 8.96%）. However, NM13 （9.58% vs. 12.02%） showed greater enhancement than XM33 （10.25% vs. 11.7%） at 28 DPA. The protein content in the dorsal and abdominal endosperms of the two wheat cultivars decreased at 12 DPA and then increased until 38 DPA. Drought stress significantly increased the protein contents in the two main regions. From 12 to 38 DPA, the amount of PB accumulation and the protein content were higher in XM33 than those in NM13. The results revealed that PB distribution varied in different endosperm tissues and that the amount of PB accumulation was remarkably augmented by drought stress.展开更多
Fasudil,a Rho-associated protein kinase(ROCK)inhibitor,has a protective effect on the central nervous system.In addition,environmental enrichment is a promising technique for inducing the recovery of motor impairments...Fasudil,a Rho-associated protein kinase(ROCK)inhibitor,has a protective effect on the central nervous system.In addition,environmental enrichment is a promising technique for inducing the recovery of motor impairments in ischemic stroke models.The present study aimed to explore whether environmental enrichment combined with fasudil can facilitate motor function recovery and induce cortical axonal regeneration after stroke.First,a mouse model of ischemic cerebral stroke was established by photochemical embolization of the left sensorimotor cortex.Fasudil solution(10 mg/kg per day)was injected intraperitoneally for 21 days after the photothrombotic stroke.An environmental enrichment intervention was performed on days 7-21 after the photothrombotic stroke.The results revealed that environmental enrichment combined with fasudil improved motor function,increased growth-associated protein 43 expression in the infarcted cerebral cortex,promoted axonal regeneration on the contralateral side,and downregulated ROCK,p-LIM domain kinase(LIMK)1,and p-cofilin expression.The combined intervention was superior to monotherapy.These findings suggest that environmental enrichment combined with fasudil treatment promotes motor recovery after stroke,at least partly by stimulating axonal regeneration.The underlying mechanism might involve ROCK/LIMK1/cofilin pathway regulation.This study was approved by the Institutional Animal Care and Use Committee of Fudan University,China(approval No.20160858A232)on February 24,2016.展开更多
The urea denatured recombinant human granulocyte colony-stimulating factor (rhG- CSF) which was expressed in Escheriachia coli (E. coli) was refolded with simultaneous purification by strong anion exchange chromatogra...The urea denatured recombinant human granulocyte colony-stimulating factor (rhG- CSF) which was expressed in Escheriachia coli (E. coli) was refolded with simultaneous purification by strong anion exchange chromatography (SAX) in the presence of low concentration- of urea. The effect of urea concentration on this refolding process was investigated. The obtained refolded rhG-CSF has a high specific activity of 2.3×108 U/mg, demonstrating that the proteins were completely refolded during the chromatographic process. With only one step by SAX in 40 min, purity and mass recovery of the refolded and purified rhG-CSF were 97% and 43%, respectively.展开更多
Olive (Olea europaea L.) tree is one of the most extensive and important agricultural crop in Mediterranean countries due to its beneficial health and nutritional properties and its high economic value. Currently, oli...Olive (Olea europaea L.) tree is one of the most extensive and important agricultural crop in Mediterranean countries due to its beneficial health and nutritional properties and its high economic value. Currently, olive tree constitutes the sixth most important cultivated plant in the world, spreading from the Mediterranean region of origin to new production areas such as Australia, South and North America and South Africa. However, the mobilization processes of storage materials i.e. reserve proteins during seed germination, which are largely involved in essential physiological process including plant growth and development, remain poorly understood. Morphometric and immunohistochemistry analyses of protein bodies contained in olive seed storage tissues, cotyledon and endosperm, were performed by using different microscopy techniques, including light (bright-field and fluorescence) microscopy and transmission electron microscopy. Furthermore, we used legumin-like proteins (11S-type globulins) as a molecular marker to study the mobilization of reserve proteins from PBs of cotyledons at germinating seedling stages by using immunofluorescence assays. Results demonstrated that cotyledon and endosperm are characterized by distinct PBs populations containing legumin-like proteins, distinctly discriminated by the number of PBs per cell and tissue, size, immunofluorescence and histochemical staining. These features reflect differential PBs biogenesis during development and maturation processes in olive seed tissues endosperm and cotyledon, in relation to proteins (polypeptides) final composition, SSPs processing and/or packaging during seed maturation. Three different mobilization patterns of legumin-like proteins were identified for the first time in cotyledon PBs during seedling germinating process. Mature proteins composition and/or processing, cell types and enzyme composition and/or differential activation have been discussed as key features determining how proteins mobilize from PBs for further degradation in the 展开更多
文摘用花生(Arachis hypogaea L.)种子的子叶组织作为模式材料,Epon 812包埋的组织进行厚切片后,按下列三步染色。第一步,PAS 反应:(1)在0.5%高碘酸(0.3%硝酸配制)中10分钟;(2)自来水洗1—2分钟,蒸馏水经过;(3)锡夫试剂中30分钟;(4)偏亚硫酸氢钠溶液三次洗涤,每次2分钟;(5)自来水洗5分钟,蒸馏水经过。第二步,苏丹黑 B 液染色:(1)70%酒精中1—2分钟;(2)新鲜配制的0.3%苏丹黑 B 液(70%酒精配制)中染色,40—60℃,30分钟至1小时;(3)70%酒精中分色1分钟;(4)自来水冼,蒸馏水经过。第三步,考马斯蓝染色:(1)7%醋酸中1—2分钟;(2)1%考马斯蓝(7%醋酸配制)中染色,60℃,20分钟;(3)0.1%醋酸液中分色1分钟;(4)自来水冼5分钟,蒸馏水经过;(5)自然干燥或在40℃电热温箱中烘干。干燥后的切片用甘油胶封固。经这三步染色的切片,细胞内的多糖、脂类和蛋白体同时被显示:淀粉粒及纤维素的细胞壁为樱红色,油滴为黑色,蛋白体为蓝色。制片能长期保存。
基金the National Natural Science Foundation of China,No.81860391(to GFZ)the Natural Science Foundation of Guangxi Zhuang Autonomous Region of China,No.2018GXNSFAA281144(to GFZ)the Key Research and Development Project of Guangxi Zhuang Autonomous Region of China,No.guike AB18221021(to SHZ)。
文摘Flavonoids have been reported to have therapeutic potential for spinal cord injury.Hawthorn leaves have abundant content and species of total flavonoids,and studies of the effects of the total flavonoids of hawthorn leaves on spinal cord injury have not been published in or outside China.Therefore,Sprague-Dawley rats were used to establish a spinal cord injury model by Allen's method.Rats were intraperitoneally injected with 0.2 m L of different concentrations of total flavonoids of hawthorn leaves(5,10,and 20 mg/kg)after spinal cord injury.Injections were administered once every 6 hours,three times a day,for 14 days.After treatment with various concentrations of total flavonoids of hawthorn leaves,the Basso,Beattie,and Bresnahan scores and histological staining indicated decreases in the lesion cavity and number of apoptotic cells of the injured spinal cord tissue;the morphological arrangement of the myelin sheath and nerve cells tended to be regular;and the Nissl bodies in neurons increased.The Basso,Beattie,and Bresnahan scores of treated spinal cord injury rats were increased.Western blot assays showed that the expression levels of pro-apoptotic Bax and cleaved caspase-3 were decreased,but the expression level of the anti-apoptotic Bcl-2 protein was increased.The improvement of the above physiological indicators showed a dose-dependent relationship with the concentration of total flavonoids of hawthorn leaves.The above findings confirm that total flavonoids of hawthorn leaves can reduce apoptosis and exert neuroprotective effects to promote the recovery of the motor function of rats with spinal cord injury.This study was approved by the Ethics Committee of the Guangxi Medical University of China(approval No.201810042)in October 2018.
基金supported by the National Natural Science Foundation of China (31171482,31571573)a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions,China (PAPD)+1 种基金the Foundation of Excellent Doctoral Dissertation of Yangzhou UniversityChina and the Jiangsu Province College Students Innovation Training Program in 2015,China (201511117063Y)
文摘The structural characteristics of protein body accumulation in different endosperm regions of hard wheat cultivar （XM33） and soft wheat cultivar （NM13） under drought stress were investigated. Drought stress treatment was implemented from plant regreening to the caryopsis mature stage. Microscope images of endosperm cells were obtained using resin semi- thin slice technology to observe the distribution and relative area of protein body （PB）. Compared with NM13, relative PB area of XM33 was significantly higher in sub-aleurone endosperm region. The amount of accumulation, including the size and relative area of PB, in two wheat cultivars was higher in sub-aleurone region than that in central region at 18 days post anthesis （DPA）. Drought stress significantly enhanced the sizes and relative areas of PBs in the dorsal and abdominal endosperms in two wheat cultivars. Particularly for dorsal endosperm, drought stress enhanced the relative PB area at 18 DPA and NM13 （5.0% vs. 6.73%） showed less enhancement than XM33 （5.49% vs. 8.96%）. However, NM13 （9.58% vs. 12.02%） showed greater enhancement than XM33 （10.25% vs. 11.7%） at 28 DPA. The protein content in the dorsal and abdominal endosperms of the two wheat cultivars decreased at 12 DPA and then increased until 38 DPA. Drought stress significantly increased the protein contents in the two main regions. From 12 to 38 DPA, the amount of PB accumulation and the protein content were higher in XM33 than those in NM13. The results revealed that PB distribution varied in different endosperm tissues and that the amount of PB accumulation was remarkably augmented by drought stress.
基金This study was supported by the National Natural Science Foundation of China,Nos.81672242(to YW),81972141(to YW)Shanghai Sailing Program,No.20YF1403500(to QZ)and Shanghai Municipal Key Clinical Specialty of China,No.shslczdzk02702(to YW).
文摘Fasudil,a Rho-associated protein kinase(ROCK)inhibitor,has a protective effect on the central nervous system.In addition,environmental enrichment is a promising technique for inducing the recovery of motor impairments in ischemic stroke models.The present study aimed to explore whether environmental enrichment combined with fasudil can facilitate motor function recovery and induce cortical axonal regeneration after stroke.First,a mouse model of ischemic cerebral stroke was established by photochemical embolization of the left sensorimotor cortex.Fasudil solution(10 mg/kg per day)was injected intraperitoneally for 21 days after the photothrombotic stroke.An environmental enrichment intervention was performed on days 7-21 after the photothrombotic stroke.The results revealed that environmental enrichment combined with fasudil improved motor function,increased growth-associated protein 43 expression in the infarcted cerebral cortex,promoted axonal regeneration on the contralateral side,and downregulated ROCK,p-LIM domain kinase(LIMK)1,and p-cofilin expression.The combined intervention was superior to monotherapy.These findings suggest that environmental enrichment combined with fasudil treatment promotes motor recovery after stroke,at least partly by stimulating axonal regeneration.The underlying mechanism might involve ROCK/LIMK1/cofilin pathway regulation.This study was approved by the Institutional Animal Care and Use Committee of Fudan University,China(approval No.20160858A232)on February 24,2016.
基金This work is supported by the National Natural Science Foundation of China(No．20175016)
文摘The urea denatured recombinant human granulocyte colony-stimulating factor (rhG- CSF) which was expressed in Escheriachia coli (E. coli) was refolded with simultaneous purification by strong anion exchange chromatography (SAX) in the presence of low concentration- of urea. The effect of urea concentration on this refolding process was investigated. The obtained refolded rhG-CSF has a high specific activity of 2.3×108 U/mg, demonstrating that the proteins were completely refolded during the chromatographic process. With only one step by SAX in 40 min, purity and mass recovery of the refolded and purified rhG-CSF were 97% and 43%, respectively.
文摘Olive (Olea europaea L.) tree is one of the most extensive and important agricultural crop in Mediterranean countries due to its beneficial health and nutritional properties and its high economic value. Currently, olive tree constitutes the sixth most important cultivated plant in the world, spreading from the Mediterranean region of origin to new production areas such as Australia, South and North America and South Africa. However, the mobilization processes of storage materials i.e. reserve proteins during seed germination, which are largely involved in essential physiological process including plant growth and development, remain poorly understood. Morphometric and immunohistochemistry analyses of protein bodies contained in olive seed storage tissues, cotyledon and endosperm, were performed by using different microscopy techniques, including light (bright-field and fluorescence) microscopy and transmission electron microscopy. Furthermore, we used legumin-like proteins (11S-type globulins) as a molecular marker to study the mobilization of reserve proteins from PBs of cotyledons at germinating seedling stages by using immunofluorescence assays. Results demonstrated that cotyledon and endosperm are characterized by distinct PBs populations containing legumin-like proteins, distinctly discriminated by the number of PBs per cell and tissue, size, immunofluorescence and histochemical staining. These features reflect differential PBs biogenesis during development and maturation processes in olive seed tissues endosperm and cotyledon, in relation to proteins (polypeptides) final composition, SSPs processing and/or packaging during seed maturation. Three different mobilization patterns of legumin-like proteins were identified for the first time in cotyledon PBs during seedling germinating process. Mature proteins composition and/or processing, cell types and enzyme composition and/or differential activation have been discussed as key features determining how proteins mobilize from PBs for further degradation in the
