Summary: To explore the mechanism of interleukin-1beta ( IL-1β) in the onset of seizure and the effect of IL-1β on the expression of adenylyl cyclase (AC) in rats with seizure induced by L-glutamate. Experimental ra...Summary: To explore the mechanism of interleukin-1beta ( IL-1β) in the onset of seizure and the effect of IL-1β on the expression of adenylyl cyclase (AC) in rats with seizure induced by L-glutamate. Experimental rats were first injected with IL-1β and then L-glutamate (a dose under the threshold) was injected into the right lateral ventricle. The rats were sacrificed 4 h after the onset of epileptic activity and examined for changes in behavior, immunohistochemistry and compared with those with seizure induced by L-glutamate alone. It was found that the expression of AC in hippocampal and neocortex of rats with seizure induced by IL-1β and L-glutamate were stronger than that of control group (P<0.05), without significant difference found between the L-glutamate group and IL-1β plus L-glutamate group in the expression of AC, the latent period and the severity of seizure. When IL-ra were given (i.c.v.) first, there was no epileptic activity and the expression of AC did not increase. There were no differences in the expression of AC of rats with IL-1ra and that of control rats. But when 2-methyl-2-(carboxycyclopropyl)glycine (MCCG) was given (i.c.v.) first, the strongest expression of AC, the shortest latent period and the the most serious seizure activities were observed. The results indicated that IL-1β could facilitate the onset of epilepsy induced by L-glutamate through IL-1R, metabotropic glutamate receptors might work with IL-1R and the increased expression of AC might be involved in the process.展开更多
目的:探讨腺苷酸环化酶相关蛋白1(adenylate cyclase-associated protein 1,CAP1)在胃癌组织中的表达及临床病理学意义。方法:采用免疫组织化学方法和组织芯片技术检测CAP1在胃癌病人石蜡标本蛋白质表达水平,分析其与胃癌临床病理特征...目的:探讨腺苷酸环化酶相关蛋白1(adenylate cyclase-associated protein 1,CAP1)在胃癌组织中的表达及临床病理学意义。方法:采用免疫组织化学方法和组织芯片技术检测CAP1在胃癌病人石蜡标本蛋白质表达水平,分析其与胃癌临床病理特征及预后的关系。结果:结果显示,CAP1在胃癌病人的表达率为55.7%(127/228)。CAP1的表达与胃癌的分化程度(P=0.037)、脉管内癌栓形成(P=0.006)、神经侵犯(P=0.05)、T分期(P=0.014)、N分期(P=0.007)以及TNM分期(P=0.026)相关。采用COX回归模型的多因素分析显示,CAP1表达、分化程度、T分期、N分期和M分期是病人生存率的独立不良预后因素。CAP1可作为影响胃癌预后的独立危险因素。结论:胃癌组织的CAP1表达可能与肿瘤细胞的侵袭和转移有关。CAP1表达提示预后不良。展开更多
The effect of intrathecal injection of dynorphin A (1-17) on second messenger systems of spinal cord relative to behavioral change in rats was studied. Dynorphin A (1-17) 5 ,10 (20nmol) caused dose-dependent flaccid p...The effect of intrathecal injection of dynorphin A (1-17) on second messenger systems of spinal cord relative to behavioral change in rats was studied. Dynorphin A (1-17) 5 ,10 (20nmol) caused dose-dependent flaccid paralysis of hindlimbs. Dynorphin A (1-17) 10, 20 nmol dose-dependently decreased spinal adenylate cyclase (AC) activity, cyclic AMP production, calmodulin (CaM) level and cyclic-nucleotide phosphodiesterase(PDE)activity 10 min after intrathecal injection. They recovered to a varying extent two hours later. Pretreatment with selective κ-opioid receptor antagonist nor-BNI 30 nmol 10 min before dynorphin A (1-17) markedly antagonized the effects of dynorphin A (1-17 ) at 20 nmol on hindlimb paralysis and inhibition of intracellular second messengers. The L-type calcium channel blocker verapamil (100nmol) also played a role in blocking dynorphin neurotoxicity. The NMDA receptor antagonist APV could partially or completely block dynorphin inhibition of CaM level and PDE activity without affecting paralysis and decrease of AC-cAMP level induced by dynorphin A(1-17) 10 min after intrathecal injection.展开更多
Our study investigated effects of C-type natriuretic peptide (CNP) on atrial dynamics and hypoxia inducible factor 1 alpha (HIF-1α) activity in perfused beating rat atria, under hypoxic conditions. Hypoxia significan...Our study investigated effects of C-type natriuretic peptide (CNP) on atrial dynamics and hypoxia inducible factor 1 alpha (HIF-1α) activity in perfused beating rat atria, under hypoxic conditions. Hypoxia significantly increased the levels of HIF-1α, concomitant with decreased trial dynamics. CNP (0.1 μmol/L) further decreased atrial dynamics under hypoxia and suppressed hypoxia-induced stimulation of HIF-1α expression. An adenylylcyclase (AC) activator, forskolin (0.1 μmol/L), significantly up-regulated atrial phosphodiesterase subtype 3A (PDE 3A) protein without affecting hypoxia-induced dynamics. In the presence of forskolin, the inhibitory effects of CNP on hypoxia-induced atrial dynamics and HIF-1α levels were significantly attenuated. Forskolin also prevented hypoxia-induced downregulation of PDE3A protein. These findings suggested that CNP inhibited atrial dynamics and HIF-1α activity in the isolated perfused beating rat atria under hypoxic conditions. Furthermore, both effects were modulated by the AC activator forskolin, through activation of CNP-PDE 3A signaling.展开更多
A preliminary study on the interaction of G protein (guanine triphosphate binding pro- tein) b1g2 subunits and their coupled components in cell signal transduction was conducted in vitro. The insect cell lines, Sf9 (S...A preliminary study on the interaction of G protein (guanine triphosphate binding pro- tein) b1g2 subunits and their coupled components in cell signal transduction was conducted in vitro. The insect cell lines, Sf9 (Spodoptera frugiperda) and H5 (Trichoplusia ni ) were used to express the recombinant protein Gb1g2. The cell membrane containing Gb1g2 was isolated through affinity chromatography column with Ni-NTA agarose by FPLC method, and the highly purified protein was obtained. The adenylyl cyclase 2 (AC2) activity assay showed that the purified Gb1g2 could signifi-cantly stimulate AC2 activity. The interaction of b1g2 subunits of G protein with the cytoplasmic tail of various mammalian adenylyl cyclases was monitored by BIAcore technology using NTA sensor chip, which relies on the phenomenon of surface plasmon resonance (SPR). The experiments showed the direct binding of Gb1g2 to the cytoplasmic tail C2 domain of AC2. The specific binding domain of AC2 with Gb1g2 was the same as AC2 activity domain which was stimulated by Gb1g2.展开更多
Tocopherols synthesized exclusively by photosynthetic organisms are major antioxidants in biomembranes.In plants,tocopherol cyclase(TC/VTE1) catalyzes the conversion of 2,3-dimethyl-5-phytyl-1,4-benzoquinone(DMPBQ) to...Tocopherols synthesized exclusively by photosynthetic organisms are major antioxidants in biomembranes.In plants,tocopherol cyclase(TC/VTE1) catalyzes the conversion of 2,3-dimethyl-5-phytyl-1,4-benzoquinone(DMPBQ) to γ-tocopherol.In the present study,OsVTE1,which encodes a rice tocopherol cyclase ortholog,was cloned and characterized.OsVTE1 was induced significantly by abiotic stresses such as high salt,H2O2,drought,cold and by the plant hormones ABA and salicylic acid.The tissue-specific expression pattern and OsVTE1-promoter GUS activity assay showed that OsVTE1 was mainly expressed in the leaf,and also could be detected in the root,stem and panicle.Compared with control plants,transgenic plants with OsVTE1 RNA interference(OsVTE1-RNAi) were more sensitive to salt stress whereas,in contrast,transgenic plants overexpressing OsVTE1(OsVTE1-OX) showed higher tolerance to salt stress.The DAB in vivo staining showed that OsVTE1-OX plants accumulated less H2O2 than did control plants.展开更多
文摘Summary: To explore the mechanism of interleukin-1beta ( IL-1β) in the onset of seizure and the effect of IL-1β on the expression of adenylyl cyclase (AC) in rats with seizure induced by L-glutamate. Experimental rats were first injected with IL-1β and then L-glutamate (a dose under the threshold) was injected into the right lateral ventricle. The rats were sacrificed 4 h after the onset of epileptic activity and examined for changes in behavior, immunohistochemistry and compared with those with seizure induced by L-glutamate alone. It was found that the expression of AC in hippocampal and neocortex of rats with seizure induced by IL-1β and L-glutamate were stronger than that of control group (P<0.05), without significant difference found between the L-glutamate group and IL-1β plus L-glutamate group in the expression of AC, the latent period and the severity of seizure. When IL-ra were given (i.c.v.) first, there was no epileptic activity and the expression of AC did not increase. There were no differences in the expression of AC of rats with IL-1ra and that of control rats. But when 2-methyl-2-(carboxycyclopropyl)glycine (MCCG) was given (i.c.v.) first, the strongest expression of AC, the shortest latent period and the the most serious seizure activities were observed. The results indicated that IL-1β could facilitate the onset of epilepsy induced by L-glutamate through IL-1R, metabotropic glutamate receptors might work with IL-1R and the increased expression of AC might be involved in the process.
文摘目的:探讨腺苷酸环化酶相关蛋白1(adenylate cyclase-associated protein 1,CAP1)在胃癌组织中的表达及临床病理学意义。方法:采用免疫组织化学方法和组织芯片技术检测CAP1在胃癌病人石蜡标本蛋白质表达水平,分析其与胃癌临床病理特征及预后的关系。结果:结果显示,CAP1在胃癌病人的表达率为55.7%(127/228)。CAP1的表达与胃癌的分化程度(P=0.037)、脉管内癌栓形成(P=0.006)、神经侵犯(P=0.05)、T分期(P=0.014)、N分期(P=0.007)以及TNM分期(P=0.026)相关。采用COX回归模型的多因素分析显示,CAP1表达、分化程度、T分期、N分期和M分期是病人生存率的独立不良预后因素。CAP1可作为影响胃癌预后的独立危险因素。结论:胃癌组织的CAP1表达可能与肿瘤细胞的侵袭和转移有关。CAP1表达提示预后不良。
文摘The effect of intrathecal injection of dynorphin A (1-17) on second messenger systems of spinal cord relative to behavioral change in rats was studied. Dynorphin A (1-17) 5 ,10 (20nmol) caused dose-dependent flaccid paralysis of hindlimbs. Dynorphin A (1-17) 10, 20 nmol dose-dependently decreased spinal adenylate cyclase (AC) activity, cyclic AMP production, calmodulin (CaM) level and cyclic-nucleotide phosphodiesterase(PDE)activity 10 min after intrathecal injection. They recovered to a varying extent two hours later. Pretreatment with selective κ-opioid receptor antagonist nor-BNI 30 nmol 10 min before dynorphin A (1-17) markedly antagonized the effects of dynorphin A (1-17 ) at 20 nmol on hindlimb paralysis and inhibition of intracellular second messengers. The L-type calcium channel blocker verapamil (100nmol) also played a role in blocking dynorphin neurotoxicity. The NMDA receptor antagonist APV could partially or completely block dynorphin inhibition of CaM level and PDE activity without affecting paralysis and decrease of AC-cAMP level induced by dynorphin A(1-17) 10 min after intrathecal injection.
文摘Our study investigated effects of C-type natriuretic peptide (CNP) on atrial dynamics and hypoxia inducible factor 1 alpha (HIF-1α) activity in perfused beating rat atria, under hypoxic conditions. Hypoxia significantly increased the levels of HIF-1α, concomitant with decreased trial dynamics. CNP (0.1 μmol/L) further decreased atrial dynamics under hypoxia and suppressed hypoxia-induced stimulation of HIF-1α expression. An adenylylcyclase (AC) activator, forskolin (0.1 μmol/L), significantly up-regulated atrial phosphodiesterase subtype 3A (PDE 3A) protein without affecting hypoxia-induced dynamics. In the presence of forskolin, the inhibitory effects of CNP on hypoxia-induced atrial dynamics and HIF-1α levels were significantly attenuated. Forskolin also prevented hypoxia-induced downregulation of PDE3A protein. These findings suggested that CNP inhibited atrial dynamics and HIF-1α activity in the isolated perfused beating rat atria under hypoxic conditions. Furthermore, both effects were modulated by the AC activator forskolin, through activation of CNP-PDE 3A signaling.
基金This work was sup-ported by the National Natural Science Foundation of China (Grant No. 30170615) and National Major Basis Study Develop-mental Plan 973 Project (TG2000016208).
文摘A preliminary study on the interaction of G protein (guanine triphosphate binding pro- tein) b1g2 subunits and their coupled components in cell signal transduction was conducted in vitro. The insect cell lines, Sf9 (Spodoptera frugiperda) and H5 (Trichoplusia ni ) were used to express the recombinant protein Gb1g2. The cell membrane containing Gb1g2 was isolated through affinity chromatography column with Ni-NTA agarose by FPLC method, and the highly purified protein was obtained. The adenylyl cyclase 2 (AC2) activity assay showed that the purified Gb1g2 could signifi-cantly stimulate AC2 activity. The interaction of b1g2 subunits of G protein with the cytoplasmic tail of various mammalian adenylyl cyclases was monitored by BIAcore technology using NTA sensor chip, which relies on the phenomenon of surface plasmon resonance (SPR). The experiments showed the direct binding of Gb1g2 to the cytoplasmic tail C2 domain of AC2. The specific binding domain of AC2 with Gb1g2 was the same as AC2 activity domain which was stimulated by Gb1g2.
基金supported by the National Basic Research Program of China (Grant No. 2006CB100102)National High-Tech Project (Grant No. 2006AA10Z18201)
文摘Tocopherols synthesized exclusively by photosynthetic organisms are major antioxidants in biomembranes.In plants,tocopherol cyclase(TC/VTE1) catalyzes the conversion of 2,3-dimethyl-5-phytyl-1,4-benzoquinone(DMPBQ) to γ-tocopherol.In the present study,OsVTE1,which encodes a rice tocopherol cyclase ortholog,was cloned and characterized.OsVTE1 was induced significantly by abiotic stresses such as high salt,H2O2,drought,cold and by the plant hormones ABA and salicylic acid.The tissue-specific expression pattern and OsVTE1-promoter GUS activity assay showed that OsVTE1 was mainly expressed in the leaf,and also could be detected in the root,stem and panicle.Compared with control plants,transgenic plants with OsVTE1 RNA interference(OsVTE1-RNAi) were more sensitive to salt stress whereas,in contrast,transgenic plants overexpressing OsVTE1(OsVTE1-OX) showed higher tolerance to salt stress.The DAB in vivo staining showed that OsVTE1-OX plants accumulated less H2O2 than did control plants.
