BACKGROUND： Tyrosine hydroxylase and phenylethanolamine-n-methyl transferase expression coexist in Purkinje cells of the rat cerebellum. Numerous reports have also been published addressing whether dopamine-beta-hydr...BACKGROUND： Tyrosine hydroxylase and phenylethanolamine-n-methyl transferase expression coexist in Purkinje cells of the rat cerebellum. Numerous reports have also been published addressing whether dopamine-beta-hydroxylase （DBH） expression exists in cerebellar Purkinje cells. OBJECTIVE： To investigate the coexistence of DBH and activator protein-2α expression in rat cerebellar Purkinje cells. DESIGN, TIME AND SETTING： A cell morphological study was performed at the Institute of Neuroscience, Chongqing Medical University, China in May 2007. MATERIALS： Ten healthy Wistar rats, of either gender, aged 14 weeks, served as experimental animals. Rabbit anti-mouse DBH, goat anti-mouse activator protein-2α and rabbit anti-mouse β-actin （Santa Cruz Biotechnology, Inc., USA）, horseradish peroxidase-labeled goat anti-rabbit IgG, FITC-labeled mouse anti-rabbit IgG, and Cy3-labeled mouse anti-goat IgG （Boster, Wuhan, China）, were used in this study. METHODS： Immunohistochemical staining was used to measure the expression of DBH or activator protein-2α, with double-label immunofluorescence being employed to determine coexpression of both, in the cerebellum of 5 randomly selected rats. Western blot assay was utilized to determine the expression of DBH and activator protein-2α in the cerebellum of the remaining 5 rats. MAIN OUTCOME MEASURES： Expression, localization and coexistence of DBH and activator protein-2α in the cerebellum were measured separately. RESULTS： Immunohistochemical staining demonstrated that cerebellar Purkinje cells stained positive for DBH and activator protein-2α. Western blot assay also demonstrated DBH and activator protein-2α expression in the cerebellum. Double-labeling immunofluorescence showed the coexistence of DBH and activator protein-2α in cerebellar Purkinje cells. CONCLUSION： Norepinephrine and activator protein-2α coexist in rat cerebellar Purkinje cells.展开更多
Puerarin, a major isoflavonoid derived from the Chinese medical herb radix puerariae （Gegen）, has been reported to inhibit neuronal apoptosis and play an anti-inflammatory role in focal cerebral ischemia model rats....Puerarin, a major isoflavonoid derived from the Chinese medical herb radix puerariae （Gegen）, has been reported to inhibit neuronal apoptosis and play an anti-inflammatory role in focal cerebral ischemia model rats. Recent findings regarding stroke pathophysiology have recognized that anti-inflammation is an important target for the treatment of ischemic stroke. The cholinergic anti-inflammatory pathway is a highly robust neural-immune mechanism for inflammation control. This study was to investigate whether activating the cholinergic anti-inflammatory pathway can be involved in the mechanism of inhibiting the inflammatory response during puerarin-induced cerebral ischemia/reperfusion in rats. Results showed that puerarin pretreatment （intravenous injection） re- duced the ischemic infarct volume, improved neurological deficit after cerebral ischemia/reperfusion and decreased the levels of interleukin-1β, interleukin-6 and tumor necrosis factor-a in brain tissue. Pretreatment with puerarin （intravenous injection） attenuated the inflammatory response in rats, which was accompanied by janus-activated kinase 2 （JAK2） and signal transducers and activators of transcription 3 （STAT3） activation and nuclear factor kappa B （NF-KB） inhibition. These observa- tions were inhibited by the alpha7 nicotinic acetylcholine receptor （a7nAchR） antagonist a-bungarotoxin （a-BGT）. In addition, puerarin pretreatment increased the expression of a7nAchR mRNA in ischemic cerebral tissue. These data demonstrate that puerarin pretreatment strongly protects the brain against cerebral ischemia/reperfusion injury and inhibits the inflammatory re- sponse. Our results also indicated that the anti-inflammatory effect of puerarin may partly be medi- ated through the activation of the cholinergic anti-inflammatory pathway.展开更多
Objective: To evaluate the effect of short-term and long-term immunization of recombinant disorganized muscle protein-1(r DIM-1) in rodents against human filarial parasite Brugia malayi.Methods: Recombinant Brugia mal...Objective: To evaluate the effect of short-term and long-term immunization of recombinant disorganized muscle protein-1(r DIM-1) in rodents against human filarial parasite Brugia malayi.Methods: Recombinant Brugia malayi DIM-1(rDIM-1 bm) protein was cloned, expressed and purified using a Ni-NTA affinity column. Mastomys coucha were immunized with rDIM-1 bm in three immunization schedules: short-term(3-dose of rDIM-1 bm), and long-term(booster doses till 3-and 6-week) and subsequently challenged with infective third-stage larvae of filarial parasite Brugia malayi(L3). Microfilaraemia was monitored in L3 exposed groups on day 90 post larval inoculation(p.l.i.) and continued till day 205 p.l.i. On day 205 p.l.i. all the infected animals were killed and total worm burden was estimated. Cellular proliferative response, macrophage activity, nitric oxide(NO) release, specific IgG and its subtypes, IgE, IgA and Th1(IFN-γ, TNF-ααand IL-2) and Th2(IL-4, IL-5, IL-6, IL-10 and IL-13) cytokine release were determined. Results: Of the 3 different immunization schedules, shortterm immunization(3-dose schedule) showed better reduction in microfilarial burden(36%-63%) in the peripheral circulation, adult worm load(52%), whereas long-term immunization(3-and 6-week schedule) exerted less effect on peripheral microfilariae count(9%-58%), and adult worm burden(9%-12.5%). Short-term immunization resulted in upregulation of cellular proliferation, macrophages activity, NO release, specific IgG, IgG1, IgG2 a, Ig G2 b, IgE and IgA levels and both Th1(IFN-γ, TNF-α and IL-2) and Th2(IL-4, IL-5, IL-6, IL-10 and IL-13) cytokine release whereas long-term immunization(3-and 6-week schedule) exerted less effect on parasite burden and showed mixed immunological responses. None of the rDIM-1 bm administration schedules induced any pathology in lymphoid tissues, or alteration in mast cell number and granularity. Conclusions: The short-term immunization with rDIM-1 bm(3-dose schedule) induces robust immune responses and protects the host from filarial parasite infection.展开更多
There is a growing interest in the diagnosis and treatment of patients with dementia and cognitive impairment at an early stage. Recent imaging studies have explored neural mechanisms underlying cognitive dysfunction ...There is a growing interest in the diagnosis and treatment of patients with dementia and cognitive impairment at an early stage. Recent imaging studies have explored neural mechanisms underlying cognitive dysfunction based on brain network architecture and functioning. The dorsal anterior cingulate cortex (dACC) is thought to regulate large-scale intrinsic brain networks, and plays a primary role in cognitive processing with the anterior insular cortex (aIC), thus providing salience functions. Although neural mechanisms have been elucidated at the connectivity level by imaging studies, their understanding at the activity level still remains unclear because of limited time-based resolution of conventional imaging techniques. In this study, we investigated temporal activity of the dACC during word (verb) generation tasks based on our newly developed event-related deep brain activity (ER-DBA) method using occipital electroencephalogram (EEG) alpha-2 powers with a time resolution of a few hundred milliseconds. The dACC exhibited dip-like temporal waveforms indicating deactivation in an initial stage of each trial when appropriate verbs were successfully generated. By contrast, monotonous increase was observed for incorrect responses and a decrease was detected for no responses. The dip depth was correlated with the percentage of success. Additionally, the dip depth linearly increased with increasing slow component of the DBA index at rest across all subjects. These findings suggest that dACC deactivation is essential for cognitive processing, whereas its activation is required for goal-oriented behavioral outputs, such as cued speech. Such dACC functioning, represented by the dip depth, is supported by the activity of the upper brainstem region including monoaminergic neural systems.展开更多
[Objectives] To explore the protective effect and possible mechanisms of the coenzyme Q10( CoQ 10) on the human umbilical vein endothelial cell( HUVEC) injury induced by the oxidized low-density lipoprotein( ox-LDL). ...[Objectives] To explore the protective effect and possible mechanisms of the coenzyme Q10( CoQ 10) on the human umbilical vein endothelial cell( HUVEC) injury induced by the oxidized low-density lipoprotein( ox-LDL). [Methods]With the human umbilical vein endothelial cells( HUVECs) cultured in vitro as the test target,the HUVECs were randomly divided into 5 groups: normal control group; model group; low concentration CoQ10 group( 12. 5); medium concentration CoQ10 group( 25); high concentration CoQ10 group( 50). The CCK-8 method was used to test the cell viability,and the drug concentration was screened in 60 μM of CoQ 10 toxic concentration; the total protein was extracted and Western blot was used to detect the protein expression of c-fos,c-jun and PON2; the RT-PCR method was used for determination of the content of c-fos,c-jun and PON2 mRNA. [Results]Compared with the normal group,the cell viability was significantly reduced in the 35 μg/m L high ox-LDL model group,and the cell injury was induced; compared with model group,after 12 h pre-protection,12. 5 μM,25 μM,50 μM CoQ10 could all significantly improve the survival of injured cells( P < 0. 05); compared with the normal group;the protein and mRNA levels of c-fos,c-jun,PON2 in the model group all declined; compared with the model group,the protein level and mRNA expression of c-fos,c-jun,PON2 in different CoQ10 groups increased to varying degrees( P < 0. 05). [Conclusions] CoQ 10 could reduce the HUVEC injury induced by high ox-LDL,and possible mechanism was achieved by upregulating the expression of AP-1 and PON2.展开更多
基金the National Natural Science Foundation of China.No.30270437
文摘BACKGROUND： Tyrosine hydroxylase and phenylethanolamine-n-methyl transferase expression coexist in Purkinje cells of the rat cerebellum. Numerous reports have also been published addressing whether dopamine-beta-hydroxylase （DBH） expression exists in cerebellar Purkinje cells. OBJECTIVE： To investigate the coexistence of DBH and activator protein-2α expression in rat cerebellar Purkinje cells. DESIGN, TIME AND SETTING： A cell morphological study was performed at the Institute of Neuroscience, Chongqing Medical University, China in May 2007. MATERIALS： Ten healthy Wistar rats, of either gender, aged 14 weeks, served as experimental animals. Rabbit anti-mouse DBH, goat anti-mouse activator protein-2α and rabbit anti-mouse β-actin （Santa Cruz Biotechnology, Inc., USA）, horseradish peroxidase-labeled goat anti-rabbit IgG, FITC-labeled mouse anti-rabbit IgG, and Cy3-labeled mouse anti-goat IgG （Boster, Wuhan, China）, were used in this study. METHODS： Immunohistochemical staining was used to measure the expression of DBH or activator protein-2α, with double-label immunofluorescence being employed to determine coexpression of both, in the cerebellum of 5 randomly selected rats. Western blot assay was utilized to determine the expression of DBH and activator protein-2α in the cerebellum of the remaining 5 rats. MAIN OUTCOME MEASURES： Expression, localization and coexistence of DBH and activator protein-2α in the cerebellum were measured separately. RESULTS： Immunohistochemical staining demonstrated that cerebellar Purkinje cells stained positive for DBH and activator protein-2α. Western blot assay also demonstrated DBH and activator protein-2α expression in the cerebellum. Double-labeling immunofluorescence showed the coexistence of DBH and activator protein-2α in cerebellar Purkinje cells. CONCLUSION： Norepinephrine and activator protein-2α coexist in rat cerebellar Purkinje cells.
基金supported by the Young Scientists Foundation of Hubei Provincial Health Department,No.QJX2012-16
文摘Puerarin, a major isoflavonoid derived from the Chinese medical herb radix puerariae （Gegen）, has been reported to inhibit neuronal apoptosis and play an anti-inflammatory role in focal cerebral ischemia model rats. Recent findings regarding stroke pathophysiology have recognized that anti-inflammation is an important target for the treatment of ischemic stroke. The cholinergic anti-inflammatory pathway is a highly robust neural-immune mechanism for inflammation control. This study was to investigate whether activating the cholinergic anti-inflammatory pathway can be involved in the mechanism of inhibiting the inflammatory response during puerarin-induced cerebral ischemia/reperfusion in rats. Results showed that puerarin pretreatment （intravenous injection） re- duced the ischemic infarct volume, improved neurological deficit after cerebral ischemia/reperfusion and decreased the levels of interleukin-1β, interleukin-6 and tumor necrosis factor-a in brain tissue. Pretreatment with puerarin （intravenous injection） attenuated the inflammatory response in rats, which was accompanied by janus-activated kinase 2 （JAK2） and signal transducers and activators of transcription 3 （STAT3） activation and nuclear factor kappa B （NF-KB） inhibition. These observa- tions were inhibited by the alpha7 nicotinic acetylcholine receptor （a7nAchR） antagonist a-bungarotoxin （a-BGT）. In addition, puerarin pretreatment increased the expression of a7nAchR mRNA in ischemic cerebral tissue. These data demonstrate that puerarin pretreatment strongly protects the brain against cerebral ischemia/reperfusion injury and inhibits the inflammatory re- sponse. Our results also indicated that the anti-inflammatory effect of puerarin may partly be medi- ated through the activation of the cholinergic anti-inflammatory pathway.
基金supported by Indian council of Medical Research,New Delhi,India(ICMR approval no.F/802/2010-ECD-11)CSIR,New Delhi,India,for award of Emeritus Scientist(scheme No.21(0963)/13/EMRII grant,29-10-2014)to P.K.M.
文摘Objective: To evaluate the effect of short-term and long-term immunization of recombinant disorganized muscle protein-1(r DIM-1) in rodents against human filarial parasite Brugia malayi.Methods: Recombinant Brugia malayi DIM-1(rDIM-1 bm) protein was cloned, expressed and purified using a Ni-NTA affinity column. Mastomys coucha were immunized with rDIM-1 bm in three immunization schedules: short-term(3-dose of rDIM-1 bm), and long-term(booster doses till 3-and 6-week) and subsequently challenged with infective third-stage larvae of filarial parasite Brugia malayi(L3). Microfilaraemia was monitored in L3 exposed groups on day 90 post larval inoculation(p.l.i.) and continued till day 205 p.l.i. On day 205 p.l.i. all the infected animals were killed and total worm burden was estimated. Cellular proliferative response, macrophage activity, nitric oxide(NO) release, specific IgG and its subtypes, IgE, IgA and Th1(IFN-γ, TNF-ααand IL-2) and Th2(IL-4, IL-5, IL-6, IL-10 and IL-13) cytokine release were determined. Results: Of the 3 different immunization schedules, shortterm immunization(3-dose schedule) showed better reduction in microfilarial burden(36%-63%) in the peripheral circulation, adult worm load(52%), whereas long-term immunization(3-and 6-week schedule) exerted less effect on peripheral microfilariae count(9%-58%), and adult worm burden(9%-12.5%). Short-term immunization resulted in upregulation of cellular proliferation, macrophages activity, NO release, specific IgG, IgG1, IgG2 a, Ig G2 b, IgE and IgA levels and both Th1(IFN-γ, TNF-α and IL-2) and Th2(IL-4, IL-5, IL-6, IL-10 and IL-13) cytokine release whereas long-term immunization(3-and 6-week schedule) exerted less effect on parasite burden and showed mixed immunological responses. None of the rDIM-1 bm administration schedules induced any pathology in lymphoid tissues, or alteration in mast cell number and granularity. Conclusions: The short-term immunization with rDIM-1 bm(3-dose schedule) induces robust immune responses and protects the host from filarial parasite infection.
文摘There is a growing interest in the diagnosis and treatment of patients with dementia and cognitive impairment at an early stage. Recent imaging studies have explored neural mechanisms underlying cognitive dysfunction based on brain network architecture and functioning. The dorsal anterior cingulate cortex (dACC) is thought to regulate large-scale intrinsic brain networks, and plays a primary role in cognitive processing with the anterior insular cortex (aIC), thus providing salience functions. Although neural mechanisms have been elucidated at the connectivity level by imaging studies, their understanding at the activity level still remains unclear because of limited time-based resolution of conventional imaging techniques. In this study, we investigated temporal activity of the dACC during word (verb) generation tasks based on our newly developed event-related deep brain activity (ER-DBA) method using occipital electroencephalogram (EEG) alpha-2 powers with a time resolution of a few hundred milliseconds. The dACC exhibited dip-like temporal waveforms indicating deactivation in an initial stage of each trial when appropriate verbs were successfully generated. By contrast, monotonous increase was observed for incorrect responses and a decrease was detected for no responses. The dip depth was correlated with the percentage of success. Additionally, the dip depth linearly increased with increasing slow component of the DBA index at rest across all subjects. These findings suggest that dACC deactivation is essential for cognitive processing, whereas its activation is required for goal-oriented behavioral outputs, such as cued speech. Such dACC functioning, represented by the dip depth, is supported by the activity of the upper brainstem region including monoaminergic neural systems.
基金Supported by Project of National Natural Science Foundation(81274126)
文摘[Objectives] To explore the protective effect and possible mechanisms of the coenzyme Q10( CoQ 10) on the human umbilical vein endothelial cell( HUVEC) injury induced by the oxidized low-density lipoprotein( ox-LDL). [Methods]With the human umbilical vein endothelial cells( HUVECs) cultured in vitro as the test target,the HUVECs were randomly divided into 5 groups: normal control group; model group; low concentration CoQ10 group( 12. 5); medium concentration CoQ10 group( 25); high concentration CoQ10 group( 50). The CCK-8 method was used to test the cell viability,and the drug concentration was screened in 60 μM of CoQ 10 toxic concentration; the total protein was extracted and Western blot was used to detect the protein expression of c-fos,c-jun and PON2; the RT-PCR method was used for determination of the content of c-fos,c-jun and PON2 mRNA. [Results]Compared with the normal group,the cell viability was significantly reduced in the 35 μg/m L high ox-LDL model group,and the cell injury was induced; compared with model group,after 12 h pre-protection,12. 5 μM,25 μM,50 μM CoQ10 could all significantly improve the survival of injured cells( P < 0. 05); compared with the normal group;the protein and mRNA levels of c-fos,c-jun,PON2 in the model group all declined; compared with the model group,the protein level and mRNA expression of c-fos,c-jun,PON2 in different CoQ10 groups increased to varying degrees( P < 0. 05). [Conclusions] CoQ 10 could reduce the HUVEC injury induced by high ox-LDL,and possible mechanism was achieved by upregulating the expression of AP-1 and PON2.
