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
In this study, biotinylated dextran amine （BDA） was microinjected into the left cortical motor area of the canine brain. Fluorescence microscopy results showed that a large amount of BDA-labeled pyramidal cells were...In this study, biotinylated dextran amine （BDA） was microinjected into the left cortical motor area of the canine brain. Fluorescence microscopy results showed that a large amount of BDA-labeled pyramidal cells were visible in the left cortical motor area after injection. In the left medulla oblongata, the BDA-labeled corticospinal tract was evenly distributed, with green fluorescence that had a clear boundary with the surrounding tissue. The BDA-positive corticospinal tract entered into the right lateral funiculus of the spinal cord and descended into the posterior part of the right lateral funiculus, close to the posterior horn, from cervical to sacral segments. There was a small amount of green fluorescence in the sacral segment. The distribution of BDA labeling in the canine central nervous system was consistent with the course of the corticospinal tract. Fluorescence labeling for BDA gradually diminished with time after injection. Our findings indicate that the BDA anterograde tracing technique can be used to visualize the localization and trajectory of the corticospinal tract in the canine central nervous system.展开更多
Unique endoperoxide moiety of artemisinin and its derivatives has been considered the functionality exhibiting highly potent antimalarial and anticancer activities.To investigate the mechanisms of their biological act...Unique endoperoxide moiety of artemisinin and its derivatives has been considered the functionality exhibiting highly potent antimalarial and anticancer activities.To investigate the mechanisms of their biological actions,development of suitable molecular probes including biotinylated derivatives is of extreme significance.The synthesis and preliminary biological assessment of four new biotinylated artemisinin derivatives have been reported in this work.展开更多
We investigated regeneration of the corticospinal tract and rearrangement of corticospinal nerves after spinal cord injury by biotinylated dextran amine （BDA） nerve tract tracing after neural stem cell transplantati...We investigated regeneration of the corticospinal tract and rearrangement of corticospinal nerves after spinal cord injury by biotinylated dextran amine （BDA） nerve tract tracing after neural stem cell transplantation. Neural stem cell transplantation increased motor function scores of rats at 3 weeks after spinal cord transection injury at the thoracic 10 segment. A proportion of BDA-labeled corticospinal tract regenerated through the spinal cord injury site at 12 weeks after transplantation. Electron microscopy revealed that the regenerated BDA-labeled nerve terminals formed new synaptic connections with neurons at the distal end of the injured site. These findings indicate that BDA nerve tract tracing effectively provides anatomic and morphological evidence of recovery after spinal cord injury.展开更多
BACKGROUND： The corticospinal tract is the core structure of cerebral control of extremity movement and plasticity, which are prerequisites for movement rehabilitation after brain injury. The measurement and assessme...BACKGROUND： The corticospinal tract is the core structure of cerebral control of extremity movement and plasticity, which are prerequisites for movement rehabilitation after brain injury. The measurement and assessment of plasticity changes within the corticospinal tract has become one of the key goals in this field. OBJECTIVE： To explore the effects of biotinylated dextran amine （BDA） as a neural tracer in the rat corticospinal tract and the possibilities of assessing plasticity within the corticospinal tract. DESIGN： An observational experiment. SETTING： Department of Acupuncture of Chinese Medical College, Chongqing Medical University, Department of Neurology, the Second Affiliated Hospital, Chongqing Medical University. MATERIALS： Eighteen male adult Sprague Dawley （SD） rats of clean grade, weighing 200-250 g, were provided by the experimental animal center of Chongqing Medical University. The animal procedures in this study were in accordance with the animal ethics standards. BDA was provided by Vector Laboratories Company （USA, catalogue Sp- 1140; serial number R0721 ）. METHODS. This experiment was performed in the Laboratory of Chongqing Medical University between September and December 2006. Adult SD rats were used in the experiment and 15% BDA was injected slowly with a mini-syringe through two round （3 mm diameter） holes into the left sensory and motor cortex. The center of one hole was located 3 mm anterior from the anterior fontanel and 1.5 mm left of the midline; the second hole was located 1.5 mm posterior from the anterior fontanel and 4 mm left of the midline. Three injections were made at each hole at three different levels： 1.4, 1.2, and 1 mm ventral from the surface of the flat skull. After 14 days, the brains and spinal cords were removed and frozen. Sections were cut on a cryostat and BDA transportation absorbed by axons was observed under a fluorescence microscope. MAIN OUTCOME MEASURES： Axonal absorption and transportation of BDA was observed under fluorescence展开更多
Existing visualized tracer studies of the corticospinal tract have been focused on rodents, which have markedly different spinal cord structures compared with humans. In this study, the segmental artery feeding the sp...Existing visualized tracer studies of the corticospinal tract have been focused on rodents, which have markedly different spinal cord structures compared with humans. In this study, the segmental artery feeding the spinal cord was embolized with digital subtraction angiography to establish a goat model of ischemic spinal cord injury. Biotinylated dextran amine was injected into the motor function areas of the cortex in goats with ischemic spinal cord injury. The corticospinal tract originates from the cerebral cortex motor function area, and travels towards the lateral funiculus at the contralateral spinal dorsal horn after decussation at the pyramid. The number of corticospinal tract positive fibers was found to be gradually reduced. These findings indicate that digital subtraction angiography can be applied to a goat model of ischemic spinal cord injury. Biotinylated dextran amine visualizes the course of the goat corticospinal tract in the spinal cord, which is similar to the human spinal cord. Biotinylated dextran amine is an ideal tracer for the corticospinal tract.展开更多
Chondroitin sulfate proteoglycans （CSPGs） which are produced by mature oligodendrocytes and reactive astrocytes can be upregulated after spinal cord injury and contribute to regenerative failure. Chondroitinase ABC ...Chondroitin sulfate proteoglycans （CSPGs） which are produced by mature oligodendrocytes and reactive astrocytes can be upregulated after spinal cord injury and contribute to regenerative failure. Chondroitinase ABC （ChABC） digests glycosaminoglycan chains on CSPGs and can thereby overcome CSPG-mediated inhibition. However, many current studies have used an incomplete spinal cord injury model, and examined results after 8-12 weeks of ChABC treatment. In this study, a complete rat spinal cord transection injury model was used to study the long-term effects of ChABC treatment by subarachnoid catheter. Pathology of spinal cord regeneration was compared with control 24 weeks following ChABC treatment using immunohistochemistry and axon tracing techniques. At 24 weeks after injury, neurofilament 200 expression was significantly greater in the ChABC treatment group compared with the transection group. In the ChABC treatment group, axonal growth was demonstrated by a large number of biotinylated dextran amine positive axons caudal to, or past, the epicenter of injury. Biotinylated dextran amine-labeled fibers were found in the proximal end of the spinal cord in the transection alone group. These results confirm that ChABC can promote axon growth, neural regeneration, and repair after spinal cord injury in rats long after the initial injury.展开更多
Brain ischemia is one of the major causes of mortality and physical disability in recent years. In order to find out an effective method to cure the disease, many researches on brain ischemia mechanism have been done....Brain ischemia is one of the major causes of mortality and physical disability in recent years. In order to find out an effective method to cure the disease, many researches on brain ischemia mechanism have been done. Calcium is an especially interesting factor in cell injury induced by brain ischemia. Much evidence has been obtained from the studies on nerve and other tissues, implying that Ca<sup>2+</sup> is a major initiator of ischemic cell injury. With ionsensitive microelectrodes, a marked reduction of Ca<sup>2+</sup> in extracellular fluid was de-展开更多
Reference strains of enterotoxigenic Escherichia coli (ETEC), non-enterotoxigenic Escherichia coli (non-ETEC), enteropathogenic Escherichia coli (EPEC), enteroinvasive Escherichia coli (EIEC), and other enteropathogen...Reference strains of enterotoxigenic Escherichia coli (ETEC), non-enterotoxigenic Escherichia coli (non-ETEC), enteropathogenic Escherichia coli (EPEC), enteroinvasive Escherichia coli (EIEC), and other enteropathogenic bacteria were used to prove the reliability of BIO-ST-DNA probe hybridization. In addition, 417 strains of E. coli isolated from children with diarrheal diseases in Shanxi Children's Hospital were examined for BIO-ST-DNA probe hybridization. In the test, BIO-ST-DNA hybridization was compared with suckling mouse assay in identifying ST-ETEC. The results obtained by both methods showed no significant difference. It was found that identification of ST-ETEC using hybridization is a simple, sensitive and more practical method.展开更多
The receptor for autocrine motility factor (AMFR) is known to be involved in the process of AMF-mediated cell migration and metastasis. This paper describes the procedures of non-radioactive in situ hybridization (ISH...The receptor for autocrine motility factor (AMFR) is known to be involved in the process of AMF-mediated cell migration and metastasis. This paper describes the procedures of non-radioactive in situ hybridization (ISH) detection of AMFR mRNA in both paraffin-embedded surgical sections and cultured cells using either biotinylated oligonucleotide probes or digoxigenin-labeled RNA probes. The results showed that the AMFR mRNA was expressed at an enhanced level in hyperplaJstic and malignant tissues of breast and prostate cancer patient surgical specimens, indicating that the elevated AMFR expression was associated with the tissue malignancy Moreover, AMFR mRNA was detected in both normal and earcinoma cells when cultured at a subconfluent density. However, AMFR expression was inhibited in confluent normal (3T3-A31 murine fibroblast and FHs738BL human bladder) cells while it continued to express in carcinoma (J82 human bladder)and metastatic (3T3-M murine fibroblast) cells irrespective of cell density This suggested a cell-cell contact downregulation of AMFR mRNA expression in normal but not in cancer cells. The ISH data obtained in this study are closely consistent with the AMFR protein expression pattern previously reported, implying that the differential expression of AMFR gene may be regulated and controlled at the transcriptional level.展开更多
本文用生物素标记的人类1号、2号、4号染色体DNA探针进行染色体原位抑制(chromosomal in situ suppression,简称CISS)杂交以研究人和猕猴染色体的同源性。结果表明:人1号染色体与猕猴1号染色体同源。其中与猕猴lpter→lq33的同源程度高...本文用生物素标记的人类1号、2号、4号染色体DNA探针进行染色体原位抑制(chromosomal in situ suppression,简称CISS)杂交以研究人和猕猴染色体的同源性。结果表明:人1号染色体与猕猴1号染色体同源。其中与猕猴lpter→lq33的同源程度高,与猕猴lq33→lqter的同源程度相对较低;人2号染色体与猕猴13号染色体长臂、9号染色体长臂和部分短臂同源;人4号染色体与猕猴2号染色体同源。结合染色体带型比较分析,本文对人和猕猴染色体的演化关系进行了探讨,该研究进一步证明了染色体重排可能是灵长类染色体进化的主要机制。展开更多
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Fsupported by the Priority Academic Development Program of Jiangsu Higher Education Institutions
文摘In this study, biotinylated dextran amine （BDA） was microinjected into the left cortical motor area of the canine brain. Fluorescence microscopy results showed that a large amount of BDA-labeled pyramidal cells were visible in the left cortical motor area after injection. In the left medulla oblongata, the BDA-labeled corticospinal tract was evenly distributed, with green fluorescence that had a clear boundary with the surrounding tissue. The BDA-positive corticospinal tract entered into the right lateral funiculus of the spinal cord and descended into the posterior part of the right lateral funiculus, close to the posterior horn, from cervical to sacral segments. There was a small amount of green fluorescence in the sacral segment. The distribution of BDA labeling in the canine central nervous system was consistent with the course of the corticospinal tract. Fluorescence labeling for BDA gradually diminished with time after injection. Our findings indicate that the BDA anterograde tracing technique can be used to visualize the localization and trajectory of the corticospinal tract in the canine central nervous system.
基金supported by the National Natural Science Foundation of China (Grant Nos. 20672128, 90713044 and 30701026)CAS (Grant No. KSCX2-YW-R23)
文摘Unique endoperoxide moiety of artemisinin and its derivatives has been considered the functionality exhibiting highly potent antimalarial and anticancer activities.To investigate the mechanisms of their biological actions,development of suitable molecular probes including biotinylated derivatives is of extreme significance.The synthesis and preliminary biological assessment of four new biotinylated artemisinin derivatives have been reported in this work.
基金Hunan Province Science and Technology Department Plan,No.2009JT3051
文摘We investigated regeneration of the corticospinal tract and rearrangement of corticospinal nerves after spinal cord injury by biotinylated dextran amine （BDA） nerve tract tracing after neural stem cell transplantation. Neural stem cell transplantation increased motor function scores of rats at 3 weeks after spinal cord transection injury at the thoracic 10 segment. A proportion of BDA-labeled corticospinal tract regenerated through the spinal cord injury site at 12 weeks after transplantation. Electron microscopy revealed that the regenerated BDA-labeled nerve terminals formed new synaptic connections with neurons at the distal end of the injured site. These findings indicate that BDA nerve tract tracing effectively provides anatomic and morphological evidence of recovery after spinal cord injury.
基金Fund of Science and Technology Committee of Chongqing,No.2004-54-83
文摘BACKGROUND： The corticospinal tract is the core structure of cerebral control of extremity movement and plasticity, which are prerequisites for movement rehabilitation after brain injury. The measurement and assessment of plasticity changes within the corticospinal tract has become one of the key goals in this field. OBJECTIVE： To explore the effects of biotinylated dextran amine （BDA） as a neural tracer in the rat corticospinal tract and the possibilities of assessing plasticity within the corticospinal tract. DESIGN： An observational experiment. SETTING： Department of Acupuncture of Chinese Medical College, Chongqing Medical University, Department of Neurology, the Second Affiliated Hospital, Chongqing Medical University. MATERIALS： Eighteen male adult Sprague Dawley （SD） rats of clean grade, weighing 200-250 g, were provided by the experimental animal center of Chongqing Medical University. The animal procedures in this study were in accordance with the animal ethics standards. BDA was provided by Vector Laboratories Company （USA, catalogue Sp- 1140; serial number R0721 ）. METHODS. This experiment was performed in the Laboratory of Chongqing Medical University between September and December 2006. Adult SD rats were used in the experiment and 15% BDA was injected slowly with a mini-syringe through two round （3 mm diameter） holes into the left sensory and motor cortex. The center of one hole was located 3 mm anterior from the anterior fontanel and 1.5 mm left of the midline; the second hole was located 1.5 mm posterior from the anterior fontanel and 4 mm left of the midline. Three injections were made at each hole at three different levels： 1.4, 1.2, and 1 mm ventral from the surface of the flat skull. After 14 days, the brains and spinal cords were removed and frozen. Sections were cut on a cryostat and BDA transportation absorbed by axons was observed under a fluorescence microscope. MAIN OUTCOME MEASURES： Axonal absorption and transportation of BDA was observed under fluorescence
基金the National Natural Science Foundation of China, No. 30972153
文摘Existing visualized tracer studies of the corticospinal tract have been focused on rodents, which have markedly different spinal cord structures compared with humans. In this study, the segmental artery feeding the spinal cord was embolized with digital subtraction angiography to establish a goat model of ischemic spinal cord injury. Biotinylated dextran amine was injected into the motor function areas of the cortex in goats with ischemic spinal cord injury. The corticospinal tract originates from the cerebral cortex motor function area, and travels towards the lateral funiculus at the contralateral spinal dorsal horn after decussation at the pyramid. The number of corticospinal tract positive fibers was found to be gradually reduced. These findings indicate that digital subtraction angiography can be applied to a goat model of ischemic spinal cord injury. Biotinylated dextran amine visualizes the course of the goat corticospinal tract in the spinal cord, which is similar to the human spinal cord. Biotinylated dextran amine is an ideal tracer for the corticospinal tract.
基金the National Natural Science Foundation of China,No.30471759
文摘Chondroitin sulfate proteoglycans （CSPGs） which are produced by mature oligodendrocytes and reactive astrocytes can be upregulated after spinal cord injury and contribute to regenerative failure. Chondroitinase ABC （ChABC） digests glycosaminoglycan chains on CSPGs and can thereby overcome CSPG-mediated inhibition. However, many current studies have used an incomplete spinal cord injury model, and examined results after 8-12 weeks of ChABC treatment. In this study, a complete rat spinal cord transection injury model was used to study the long-term effects of ChABC treatment by subarachnoid catheter. Pathology of spinal cord regeneration was compared with control 24 weeks following ChABC treatment using immunohistochemistry and axon tracing techniques. At 24 weeks after injury, neurofilament 200 expression was significantly greater in the ChABC treatment group compared with the transection group. In the ChABC treatment group, axonal growth was demonstrated by a large number of biotinylated dextran amine positive axons caudal to, or past, the epicenter of injury. Biotinylated dextran amine-labeled fibers were found in the proximal end of the spinal cord in the transection alone group. These results confirm that ChABC can promote axon growth, neural regeneration, and repair after spinal cord injury in rats long after the initial injury.
基金Project supported by the National Natural Science Foundation of China.
文摘Brain ischemia is one of the major causes of mortality and physical disability in recent years. In order to find out an effective method to cure the disease, many researches on brain ischemia mechanism have been done. Calcium is an especially interesting factor in cell injury induced by brain ischemia. Much evidence has been obtained from the studies on nerve and other tissues, implying that Ca<sup>2+</sup> is a major initiator of ischemic cell injury. With ionsensitive microelectrodes, a marked reduction of Ca<sup>2+</sup> in extracellular fluid was de-
文摘Reference strains of enterotoxigenic Escherichia coli (ETEC), non-enterotoxigenic Escherichia coli (non-ETEC), enteropathogenic Escherichia coli (EPEC), enteroinvasive Escherichia coli (EIEC), and other enteropathogenic bacteria were used to prove the reliability of BIO-ST-DNA probe hybridization. In addition, 417 strains of E. coli isolated from children with diarrheal diseases in Shanxi Children's Hospital were examined for BIO-ST-DNA probe hybridization. In the test, BIO-ST-DNA hybridization was compared with suckling mouse assay in identifying ST-ETEC. The results obtained by both methods showed no significant difference. It was found that identification of ST-ETEC using hybridization is a simple, sensitive and more practical method.
文摘The receptor for autocrine motility factor (AMFR) is known to be involved in the process of AMF-mediated cell migration and metastasis. This paper describes the procedures of non-radioactive in situ hybridization (ISH) detection of AMFR mRNA in both paraffin-embedded surgical sections and cultured cells using either biotinylated oligonucleotide probes or digoxigenin-labeled RNA probes. The results showed that the AMFR mRNA was expressed at an enhanced level in hyperplaJstic and malignant tissues of breast and prostate cancer patient surgical specimens, indicating that the elevated AMFR expression was associated with the tissue malignancy Moreover, AMFR mRNA was detected in both normal and earcinoma cells when cultured at a subconfluent density. However, AMFR expression was inhibited in confluent normal (3T3-A31 murine fibroblast and FHs738BL human bladder) cells while it continued to express in carcinoma (J82 human bladder)and metastatic (3T3-M murine fibroblast) cells irrespective of cell density This suggested a cell-cell contact downregulation of AMFR mRNA expression in normal but not in cancer cells. The ISH data obtained in this study are closely consistent with the AMFR protein expression pattern previously reported, implying that the differential expression of AMFR gene may be regulated and controlled at the transcriptional level.
文摘本文用生物素标记的人类1号、2号、4号染色体DNA探针进行染色体原位抑制(chromosomal in situ suppression,简称CISS)杂交以研究人和猕猴染色体的同源性。结果表明:人1号染色体与猕猴1号染色体同源。其中与猕猴lpter→lq33的同源程度高,与猕猴lq33→lqter的同源程度相对较低;人2号染色体与猕猴13号染色体长臂、9号染色体长臂和部分短臂同源;人4号染色体与猕猴2号染色体同源。结合染色体带型比较分析,本文对人和猕猴染色体的演化关系进行了探讨,该研究进一步证明了染色体重排可能是灵长类染色体进化的主要机制。
