Objective To screen and identify differentially expressed genes in the dorsal root ganglion （DRG） in early experimental diabetic rats. Methods Diabetic model rats were induced by single intraperitoneal injection of ...Objective To screen and identify differentially expressed genes in the dorsal root ganglion （DRG） in early experimental diabetic rats. Methods Diabetic model rats were induced by single intraperitoneal injection of streptozotocin （STZ）. At the second week after STZ injection, the sensory nerve conduction velocities （SNCV） of sciatic nerve were measured as an indicator of neuropathy. The technique of silver-staining mRNA differential display polymerase chain reaction （DD-PCR） was used to detect the levels of differentially expressed genes in rat DRG. The cDNA fragments that displayed differentially were identified by reverse-hybridization, cloned and sequenced subsequently, and then confirmed by Northern blot. Results The SNCV in the diabetic model group [n = 9, （45.25±10.38） m/s] reduced obviously compared with the control group [n = 8, （60.10± 11.92） m/s] （P 〈 0.05）. Seven distinct cDNA clones, one was up-regulated gene and the others were downregulated ones, were isolated by silver-staining mRNA differential display method and confirmed by Northern blot. According to the results of sequence alignment with GenBank data, majority of the clones had no significant sequence similarity to previously reported genes except only one that showed high homology to 6-pyruvoyl-tetrahydropterin synthase mRNA （accession No., BC059140）, which had not been reported to relate to diabetic neuropathy. Conclusion These differentially expressed genes in the diabetic DRG may contribute to the pathogenesis of diabetic peripheral neuropathy.展开更多
Retinal ganglion cells in the rat were studied using the heavy metal intensified oytochrome oxidase and horseradish peroxidase histochemieal methods. The results show that a population of large retinal ganglion cells ...Retinal ganglion cells in the rat were studied using the heavy metal intensified oytochrome oxidase and horseradish peroxidase histochemieal methods. The results show that a population of large retinal ganglion cells was consistently observed with the eyto3hrome oxidase staining method in retinas of normal rats or rats which received unilateral thalamotomy at birrth. These oytochrome oxidase rich ganglion cells appeared to have large somata, 3-6 primary dendrites and extensive dendritic arbors, and are comparable to ganglion cells labeled by the wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). However, the morphological details of some of the cells revealed by the cytoahrome oxidase staining method are frequently better than those shown by the HRP histochemieal method. These results suggest that the mit03hondrial enzyme oytoohrome oxidase can be used as a simple but reliable marker for identifying and studying a population of retinal ganglion cells with high metabolie rate in the rat.展开更多
目的采用电生理记录和行为学观察研究了外周初级传入神经元对单独和联合使用ATP及缓激肽(BK)的反应及其机制。方法在大鼠新鲜分离背根神经节(DRG)神经元标本上应用全细胞膜片钳技术记录ATP激活电流(I_(ATP))以及BK对I_(ATP)的调制作用,...目的采用电生理记录和行为学观察研究了外周初级传入神经元对单独和联合使用ATP及缓激肽(BK)的反应及其机制。方法在大鼠新鲜分离背根神经节(DRG)神经元标本上应用全细胞膜片钳技术记录ATP激活电流(I_(ATP))以及BK对I_(ATP)的调制作用,并结合痛行为实验进行整体行为观察。结果在大鼠新鲜分离的DRG细胞上预加BK后再加ATP,则I_(ATP)明显增强,其增强程度依赖于BK的浓度(10^(-6)～10^(-4) mol/L)。预加BK后ATP的量—效曲线上移,其电流最大值与对照相比增加20.75%,而BK预加前后ATP量—效曲线的EC_(50)值非常接近(1.65×10^(-5)(?) 2.0×10^(-5)mol/L)。在大鼠后肢掌底皮下分别注射BK和ATP均引起浓度依赖性的痛行为(抬腿)反应.当联合应用BK(10^(-6)mol/L)和ATP(10^(-5),10^(-4) and 10^(-3)mol/L)时,后爪抬腿时间随ATP浓度的增大而急剧地增强。结论炎性介质BK、ATP等在外周感觉神经来梢疼痛信息的产生、传递和调制方面起着重要的作用。ATP和BK具有协同作用,这种作用是非竞争性的,BK能明显增强I_(ATP)预加BK后,随着ATP浓度的增高所诱导的痛行为反应急剧增加。展开更多
Purpose： To investigate effects of neuro-immuno-modulation on wound healing by observing changes of cytokines and hypothalamic-pituitary-adrenal （HPA） axis hormones in acute stress reaction in rats with wound and c...Purpose： To investigate effects of neuro-immuno-modulation on wound healing by observing changes of cytokines and hypothalamic-pituitary-adrenal （HPA） axis hormones in acute stress reaction in rats with wound and combined local radiation injury. Methods： Sixty female Wistar rats （weighting 200 ± 20 g） were randomly divided into normal control group, wound group and combined wound-local radiation （CWR） group （25 Gy local radiation post wound）, 20 rats in each group. Contents of IL-1β, IL-6 and IFN-γ and IL-4 in serum were measured and changes of adrenocorticotropic hormone （ACTH） and glucocorticoid （GC） in serum were analyzed by using enzyme-linked immunosorbent assay and radioimmunologic assay, respectively at different time points post wound and radiation. Results： （1） The level of IFN-γ, one of the Thl cell cytokines increased significantly at 14 d post CWIL which was markedly higher than that in control group and wound group. However, the level of IL-4, IL-1β and IL-6, one of the Th2 cell cytokines, did not show obvious change. （2） Ratio of Thl/Th2 （IFN-γ/IL-4） in wound group and CWR group increased significantly at 7 d after wound and radiation, which suggested that Thl/Th2 balance drifted to Thl immune response. The ratio of Thl/Th2 in wound group returned to the normal level up to 14 d after the wound and radiation, while the Thl/Th2 ratio in CWR group increased persistently and was much higher than that in control and wound groups. （3） Level of serous ACTH and CC in CWR group increased at 3 d post wound and radiation, and among them, level of CC showed statistically significant increase, which was much higher than that in control and wound groups. Conclusion： Level of serous neurohormone CC in rats increased significantly immediately after wound and radiation; while the level of IFN-γ showed significant increase only up to 14 d after wound and radiation, and the Th1/Th2 imbalance sustained till 28 d post wound and radiation. In order to reduce acute damage caused by CWR, organic immune system and nerve system showed up a marked regulate effects simultaneously and mutually. Nonetheless, the excessive stress induced by CWR causes distur- bance of immunoregulation, which is one of the key reasons for delayed wound healing in CWR.展开更多
基金the grant from Technical Program of Social Development ofNantong Municipality (No.S30043)the Natural ScienceFoundation of Nantong University (No. 05Z084)
文摘Objective To screen and identify differentially expressed genes in the dorsal root ganglion （DRG） in early experimental diabetic rats. Methods Diabetic model rats were induced by single intraperitoneal injection of streptozotocin （STZ）. At the second week after STZ injection, the sensory nerve conduction velocities （SNCV） of sciatic nerve were measured as an indicator of neuropathy. The technique of silver-staining mRNA differential display polymerase chain reaction （DD-PCR） was used to detect the levels of differentially expressed genes in rat DRG. The cDNA fragments that displayed differentially were identified by reverse-hybridization, cloned and sequenced subsequently, and then confirmed by Northern blot. Results The SNCV in the diabetic model group [n = 9, （45.25±10.38） m/s] reduced obviously compared with the control group [n = 8, （60.10± 11.92） m/s] （P 〈 0.05）. Seven distinct cDNA clones, one was up-regulated gene and the others were downregulated ones, were isolated by silver-staining mRNA differential display method and confirmed by Northern blot. According to the results of sequence alignment with GenBank data, majority of the clones had no significant sequence similarity to previously reported genes except only one that showed high homology to 6-pyruvoyl-tetrahydropterin synthase mRNA （accession No., BC059140）, which had not been reported to relate to diabetic neuropathy. Conclusion These differentially expressed genes in the diabetic DRG may contribute to the pathogenesis of diabetic peripheral neuropathy.
文摘Retinal ganglion cells in the rat were studied using the heavy metal intensified oytochrome oxidase and horseradish peroxidase histochemieal methods. The results show that a population of large retinal ganglion cells was consistently observed with the eyto3hrome oxidase staining method in retinas of normal rats or rats which received unilateral thalamotomy at birrth. These oytochrome oxidase rich ganglion cells appeared to have large somata, 3-6 primary dendrites and extensive dendritic arbors, and are comparable to ganglion cells labeled by the wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). However, the morphological details of some of the cells revealed by the cytoahrome oxidase staining method are frequently better than those shown by the HRP histochemieal method. These results suggest that the mit03hondrial enzyme oytoohrome oxidase can be used as a simple but reliable marker for identifying and studying a population of retinal ganglion cells with high metabolie rate in the rat.
文摘目的采用电生理记录和行为学观察研究了外周初级传入神经元对单独和联合使用ATP及缓激肽(BK)的反应及其机制。方法在大鼠新鲜分离背根神经节(DRG)神经元标本上应用全细胞膜片钳技术记录ATP激活电流(I_(ATP))以及BK对I_(ATP)的调制作用,并结合痛行为实验进行整体行为观察。结果在大鼠新鲜分离的DRG细胞上预加BK后再加ATP,则I_(ATP)明显增强,其增强程度依赖于BK的浓度(10^(-6)～10^(-4) mol/L)。预加BK后ATP的量—效曲线上移,其电流最大值与对照相比增加20.75%,而BK预加前后ATP量—效曲线的EC_(50)值非常接近(1.65×10^(-5)(?) 2.0×10^(-5)mol/L)。在大鼠后肢掌底皮下分别注射BK和ATP均引起浓度依赖性的痛行为(抬腿)反应.当联合应用BK(10^(-6)mol/L)和ATP(10^(-5),10^(-4) and 10^(-3)mol/L)时,后爪抬腿时间随ATP浓度的增大而急剧地增强。结论炎性介质BK、ATP等在外周感觉神经来梢疼痛信息的产生、传递和调制方面起着重要的作用。ATP和BK具有协同作用,这种作用是非竞争性的,BK能明显增强I_(ATP)预加BK后,随着ATP浓度的增高所诱导的痛行为反应急剧增加。
文摘Purpose： To investigate effects of neuro-immuno-modulation on wound healing by observing changes of cytokines and hypothalamic-pituitary-adrenal （HPA） axis hormones in acute stress reaction in rats with wound and combined local radiation injury. Methods： Sixty female Wistar rats （weighting 200 ± 20 g） were randomly divided into normal control group, wound group and combined wound-local radiation （CWR） group （25 Gy local radiation post wound）, 20 rats in each group. Contents of IL-1β, IL-6 and IFN-γ and IL-4 in serum were measured and changes of adrenocorticotropic hormone （ACTH） and glucocorticoid （GC） in serum were analyzed by using enzyme-linked immunosorbent assay and radioimmunologic assay, respectively at different time points post wound and radiation. Results： （1） The level of IFN-γ, one of the Thl cell cytokines increased significantly at 14 d post CWIL which was markedly higher than that in control group and wound group. However, the level of IL-4, IL-1β and IL-6, one of the Th2 cell cytokines, did not show obvious change. （2） Ratio of Thl/Th2 （IFN-γ/IL-4） in wound group and CWR group increased significantly at 7 d after wound and radiation, which suggested that Thl/Th2 balance drifted to Thl immune response. The ratio of Thl/Th2 in wound group returned to the normal level up to 14 d after the wound and radiation, while the Thl/Th2 ratio in CWR group increased persistently and was much higher than that in control and wound groups. （3） Level of serous ACTH and CC in CWR group increased at 3 d post wound and radiation, and among them, level of CC showed statistically significant increase, which was much higher than that in control and wound groups. Conclusion： Level of serous neurohormone CC in rats increased significantly immediately after wound and radiation; while the level of IFN-γ showed significant increase only up to 14 d after wound and radiation, and the Th1/Th2 imbalance sustained till 28 d post wound and radiation. In order to reduce acute damage caused by CWR, organic immune system and nerve system showed up a marked regulate effects simultaneously and mutually. Nonetheless, the excessive stress induced by CWR causes distur- bance of immunoregulation, which is one of the key reasons for delayed wound healing in CWR.
