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Application of Luxol Fast Blue staining in locating the corticospinal tract in adult rats

Neural regeneration Research 2006年第7期1卷 642-644页

作者： Su Liu1, Guangyu Shen1, Guangming Lü2, Xiaosong Gu3 1Department of Rehabilitation Medicine, Affiliated Hospital of nantong University, nantong 226001, jiangsu province, china 2Department of Human Anatomy, nantong University, nantong 226001, jiangsu province, china 3jiangsu key laboratory of nerve regeneration, nantong 226001, jiangsu province, china Department of Rehabilitation Medicine Hospital of Nantong University Nantong 226001 Jiangsu Province China Department of Human Anatomy Nantong University Nantong 226001 Jiangsu Province China jiangsu key laboratory of nerve regeneration Nantong 226001 Jiangsu Province China

BACKGROUND: There are many methods for myelin staining, mordant, or oil-soluble dye or the special reaction of osmic acid with lipoid is used according to different principles. The commonly used methods are classic Well staining, classic lithium carbonate-haematine staining, fast green staining, silver staining, etc. Luxol Fast Blue can brightly stain myelin sheath, and has certain specificity. The background can be very clean if there is proper differentiation, whereas Luxol Fast Blue is cheap and convenient to operate, thus it is an ideal staining reagent for routine myelin sheath. OBJECTIVE: To show the corticospinal tract of normal adult rats with Luxol Fast Blue staining method. DESIGN: A repetitive measurement design. SETTINGS: Institute of Nuerobiology, nantong University; Department of Rehabilitation Medicine, Affiliated Hospital of nantong University. MATERIALS: Six healthy adult male SD rats of clean degree, weighing averagely 300 g, were provided by the experimental animal center of nantong University. 1 g/L Luxol Fast Blue solution was provided by Sigma Company; Leica CM1900 cryostat microtome by Leica Company; Leica DMR microscope by Leica Company. METHODS: The experiment was carried out in the Staff Room of Human Anatomy, nantong University in May 2005. The rats were given intraperitoneal injection of combined anesthetic (2 mL/kg), then the chest was open for perfusing saline and phosphate buffer containing formamint via heart. Brain and spinal cord were removed after 1 hour then fixed, then changed to phosphate buffer (pH 7.4) containing 300 g/L saccharu at 4 ℃, and stayed overnight, tissue blocks at pyramid, decussation of pyramid and cervical, thoracic, lumbar and sacral segments of spinal cord were removed to prepare continuous horizontal frozen sections (30 μm) after sedimentation, the sections were dried at room temperature. The corticospinal tract of normal adult rats were shown with Luxol Fast Blue staining method, and observed under Leica DMR

关键词： Application of Luxol Fast Blue staining in locating the corticospinal tract in adult rats

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Biocompatibility studies of silk fibroin-based artificial nerve grafts in vitro and in vivo

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Progress in Natural Science:Materials International 2007年第9期17卷 1029-1034页

作者： Chen Xuemei,Yang Yumin,Wu Jian,Zhao Yahong,Ding Fei and Gu Xiaosong(jiangsu key laboratory of Neuroregeneration,nantong University,nantong 226001,china) jiangsu key laboratory of Neuroregeneration Nantong University Nantong 226001 China

Silk fibroin(SF)has been used extensively in the biomedical field including tissue engineering for the generation of artificial bones,skins or *** have previously reported on good in vitro biocompatibility of SF fibers with peripheral nerve tissues and *** the present study,we developed a novel design of the SF-based artificial nerve graft(SF graft)which was composed of a SF-nerve guidance conduit(NGC)inserted with SF *** assay was performed to determine the cytotoxicity of the SF-NGC extract fluid on the cultured L929 cells derived from an immortalized mouse fibroblast cell *** addition,this SF graft was implanted into adult rats for bridging a 10-mm long sciatic nerve *** following-up experiments at initial stage(1-4 week)of nerve regeneration including routine blood tests and histochemical investigation were conducted to evaluate the in vivo biocompatibility of the SF graft with peripheral *** results demonstrated that the SF-NGC graft was biocompatible with the surrounding tissues and cells due to its low inflammatory potential with a grade 0 under the *** guidelines and it was generally suitable to a certain degree for bridging peripheral nerve defects in virtue of supporting Schwann cell adherence,expansion and *** the SF graft is a promising alternative to classical autografts for peripheral nerve repair.

关键词： silk fibroin artificial nerve graft nerve guidance conduit biocompatibility cytotoxicity

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Achyranthes bidentata Blume extract promotes neuronal growth in cultured embryonic rat hippocampal neurons

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Progress in Natural Science:Materials International 2009年第5期19卷 549-555页

作者： Xin Tang1, Yiren Chen 1, Xiaosong Gu, Fei Ding jiangsu key laboratory of Neuroregeneration, nantong University, nantong 226001, china [a]jiangsu key laboratory of Neuroregeneration Nantong University Nantong 226001 China

We have prepared an aqueous extract of Achyranthes bidentata Blume, a commonly prescribed Chinese medicinal herb, and reported, in previous studies, that A. bidentata extract benefits nerve growth and prevents neuron apoptosis. In this study, we investigated the actions of A. bidentata extract on survival and growth of primarily cultured rat hippocampal neurons. The morphological observation revealed that neurite growth from hippocampal neurons was significantly enhanced by A. bidentata extract with similar effects to those induced by nerve growth factor (NGF), and the greatest neurite growth appeared on treatment with A. bidentata extract at 1 lg/ml for 24 h. DNA microarray analysis indicated that there were 25 upregulated genes and 47 downregulated genes exhibiting significantly differential expression in hippocampal neurons treated with A. bidentata extract at 1 lg/ml for 6 h when compared to those in untreated hippocampal neurons. Real-time quantitative RT-PCR and Western blot analysis demonstrated that the expression of growth-associated protein-43 in hippocampal neurons was upregulated at both mRNA and protein levels after treatment with A. bidentata extract, and the optimal dosage of the extract was also 1 lg/ml. These data confirm that A. bidentata extract could promote in vitro hippocampal neuronal growth in a dose-and time-dependent manner.

关键词： Achyranthes bidentata Blume Aqueous extract nerve growth factor Hippocampal neurons Neurite outgrowth Growth-associated protein-43 Gene differential expression

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nerve regeneration factor promotes nerve regeneration in rat

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Progress in Natural Science:Materials International 2006年第12期16卷 1281-1287页

作者： ZHANG Qi, LIU Mei, ZHOU Mingming and DING Fei**(key laboratory of Neuroregeneration, nantong University, nantong 226001, china ) [a]key laboratory of Neuroregeneration Nantong University Nantong China

nerve regeneration factor (NRF) extracted from an oral liquid of traditional Chinese medicine, as a nerve growth decoction has been reported by previous studies to exert effects of promoting nerve growth and preventing neuron apoptosis. For new insights into the function of NRF on primary cultured neurons, we investigated the neurite outgrowth in cultured rat dorsal root ganglion (DRG) explant treated with NRF by immunofluorescence and the gene and protein expressions of neurofilament-H(NF-H) and growth associated protein 43 (GAP43) in the cultured rat DRG neurons by real-time quantitative RT-PCR and Western blotting, respectively. In addition, we used a rat model of sciatic nerve crush to evaluate the effect of NRF on regeneration of injured sciatic nerve by a combination of walk track analysis, electrophysiological and histological assessments. The in vitro experiments indicated that NRF promoted the neurite growth of DRGs and the expression of NF-H and GAP43 at mRNA and protein levels in DRG neurons, and in vivo experiments showed that NRF improved peripheral nerve regeneration and functional recovery.

关键词： nerve regeneration factor dorsal root ganglion sciatic nerve crush injury neurofilament-H growth associated protein 43.

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Microelectronics-embedded channel bridging and signal regeneration of injured spinal cords

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Progress in Natural Science:Materials International 2009年第10期19卷 1261-1269页

作者： Zhigong Wanga,,Xiaosong Gub,Xiaoying Lü c,Zhenglin Jiangb,Wenyuan Li a,Guangming Lü b,Yufeng Wang a,Xiaoyan Shen a,Xintai Zhao a,Huiling Wangc,Zhenyu Zhang a,Hongmei Shen b,Yang Wua,Weixing Shen b,Jingyang Zhang a,Dong Chenc,Xiaoyi Maoa,Huaxiang Shenb a Institute of RF-& OE-ICs,Southeast University,Nanjing 210096,china b key laboratory of Neural regeneration of jiangsu province,nantong University,nantong 226001,china c State key laboratory of Bioelectronics,Southeast University,Nanjing 210096,china [a]Institute of RF- & OE-ICs Southeast University Nanjing 210096 China [b]key laboratory of Neural regeneration of jiangsu province Nantong University Nantong 226001 China [c]State key laboratory of Bioelectronics Southeast University Nanjing 210096 China

Due to the dificulty in spinal cord regeneration with biological methods,the microelectronic neural bridge,a new concept based on microelectronic technology,is *** microelectronic system has been realized in the forms of hybrid and integrated *** integrated circuits forneural signal detection,stimulation,and regeneration are realized in a CMOS *** animal experiments with 100 toads,48 rats,and 3 rabbits,nerve signals have been successfully detected from spinal cords and sciatic nerves,and functional electrical stimulation has been carried out for spinal cords and sciatic *** the microelectronic system is bridged between the controlling and stimulated nerve,the relevant motion of legs and nerve signal waveforms,which are stimulated by the evoked or spontaneous nerve signal through such a system,have been ***,the feasibility of the presented method was demonstrated.

关键词： nerve injury Microelectrode arrays Neural recordings Neurostimulation Functional recovery

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Dog sciatic nerve gap repaired by artificial tissue nerve graft

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Progress in Natural Science:Materials International 2003年第8期 20-25+83-84页

作者： GU Xiaosong, ZHANG Peiyun, WANG Xiaodong, DING Fei, PENG Luping and CHENG Hongbing(nantong Medical College, The key laboratory of Neuroregeneration of jiangsu province, nantong 226001, china) nantong Medical College The Key Laboratory of Neuroregeneration of Jiangsu Province Nantong 226001 China

The feasibility of repairing dog sciatic nerve damage by using a biodegradable artificial tissue nerve graft enriched withneuroregenerating factors is investigated. The artificial nerve graft was implanted to a 30 mm gap of the sciatic nerve damage in 7 *** dogs with the same nerve damage that were repaired by interposition of the autologous nerve or were given no treatment served as con-trol group 1 or 2, respectively. The observations include gross and morphological observations, immune reaction, electrophysiological ex-amination, fluorescence tracing of the neuron formation and the number of the neurons at the experimental sites, etc. Results showed that6 months after the implantation of the graft, the regenerated nerve repaired the damage of the sciatic nerve without occurrence of rejectionand obvious inflammatory reaction in all 7 dogs, and the function of the sciatic nerve recovered with the nerve conduction velocity of (23.91±11.35) m/s. The regenerated neurons and the forming of axon could

关键词： artificial tissue nerve medical biodegradable material peripheral nerve regeneration.

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Effect of nerve regeneration factor on differentiation of PC12 cells and its signaling pathway

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Progress in Natural Science:Materials International 2004年第11期 35-38+99页

作者： DING Fei, QIANG Liang, LIU Mei, GU Xingxing and GU Xiaosong*(The key laboratory of Neuroregeneration of jiangsu province, nantong Medical College, nantong 226001, china) ThekeylaboratoryofNeuroregenerationofjiangsuprovince NantongMedicalCollegeNantong226001China

The effects of nerve regeneration factor (NRF) on neuronal differentiation of PC12 cells and its signaling pathway are investigated by morphological observation and immunofluorescent cytochemical method, and the activity of ERK1/2 in NRF-treated PC12 cells in absence of serum is also studied by immuno-coprecipitation and Western blot analysis. The MEK1/2-specific inhibitor U0126, the broad-spectrum protein kinase C (PKC) inhibitor G6983 and tyrosine protein kinase (TPK) inhibitor genistein were used to determine the roles of the activation of ERK1/2 by NRF and the involvement of certain kinds of PKC or TPK receptor in this activation process. The results show that U0126 and G6983 inhibit the activation of ERK1/2 by NRF to different extents, while genistein has no effect on it, demonstrating that NRF remarkably induces neuronal differentiation of PC12 cells through activating ERK1/2 in a dose-dependent and time-dependent manner.

关键词： nerve regeneration factor PC12 cells neuronal differentiation ERK1/2.

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The Experimental Study on the Artificial Tissue nerve Graft in Repairing Sciatic nerve Gap in Dogs

The Experimental Study on the Artificial Tissue Nerve Graft ...

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中国组织工程、干细胞与神经再生学术会议

作者： Wen Hu Xiaodong Wang Yong Cao Jian Yao Jian Wu Xiaosong Gu* jiangsu province key laboratory of Neuroregeneration,nantong University,nantong,jiangsu 226001,P.R.china

We have developed a dual-component artificial tissue nerve graftcomprising an outer microporous conduit of chitosan and internal orientedfilaments of polyglycolic acid(PGA).The novel graft was used for bridgingsciatic nerve across a 30 mm gap in 6 beagle *** other 6 beagle dogsas the positive control were implanted with an autologous nerve instead ofthe artificial graft and the other 5 beagle dogs as the negative control wasunimplanted with sciatic nerve lesions left *** animals were monitoredfor changes in their appearance and locomotion activities after *** and gait were regularly recorded by the aid of photos and videotapesfor each *** months postoperatively,electrophysiological assessment,FluroGold retrograde tracing,histological methods including light microscopyand transmission electron microscopy,immunohistochemistry and morphometricanalyses of regenerated nerves as well as target muscles were applied toevaluate the nerve repair effects of the artificial *** morphologicalobservations together with etectrophysiological and histomorphometricalmeasurements,including the compound muscle action potential and conductionmotor nerve velocity,the percentage of neurofilament immunopositive areasand myeliated axon density,the thickness of myelin sheath and G ratio(myelinated axon diameter total myelinated fiber diameter),the wet weightratio of gastrocnemius muscle and average percentage of the collagen fiberarea,etc.,demonstrated that the sciatic nerve truck had been reconstructed,with restoration of nerve continuity and functional recovery,and the targetskeletal muscle had been reinnervated,improving locomotion activities of theoperated *** is also shown that the nerve repair effects with ourartificial graft were comparable with those with ***,thisstudy proves the feasibility of a novel artificial graft we developed for promoting nerve regeneration,and establishes an experimental basis for thedevelopment of ti

关键词： Peripheral nerve regeneration Tissue engineering Artificial nerve graft Chitosan Polyglycolic acid Long nerve gaps Dogs

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The experimental study on the transplantation of Nurrl gene transfected neural stem cells for treatment of Parkinson’s disease

The experimental study on the transplantation of Nurrl gene ...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Xue-Feng TAN,Guo-Hua JIN~*,Mei-Ling TIAN,Hao-Ming LI,Hui-Xia ZHU,Lei ZHANG Department of Anatomy and Neurobiology,the key laboratory of nerve regeneration of jiangsu province,School of Medicine,nantong University,nantong 226001,china

＜正＞Objective To study the differentiation of neural stem cells（NSCs）transfected with nuclear receptor related factor 1（Nurr1）gene into dopaminergic neurons and its behavioral exertion after transplanted into the striatum of Parkinson’s disease（PD）*** The unilateral PD rats were prepared by the rat brain stereotaxic technology using single-point injection of 6-OHDA into the right medial forebrain *** recombinant pEGFP-N1 -Nurr1 was transfected into NSCs through elctroporation,and the expression of Nurrl gene was detected by RTPCR. The NSCs were labled with DIL and transplanted into the right striatum of PD ***-induced rotation test was performed to observe the behaviour amelioration after *** the expression of tyrosine hydroxylase（TH）was detected by immunofluorescence *** PD rats appear typical rotations, and the rate of success was 62%.Nurrl gene was overexpressed after transfected with recombinant *** transplantation,the behaviour of PD rats did not ameliorate obviously in sham and NSCs *** positive cells could be observed in NSCs group,but few of them exhibited TH positive;In Nurr1 group,the rotations of PD rats began to decrease at the sixth week after transplantation,and there were many DIL/TH double-labeled neurons in transplanted *** Overexpression of Nurrl could induce the differentiation of NSCs into TH positive dopaminergic neurons in the striatum of PD rats,and ameliorate the behavioral function of PD rats.

关键词： nuclear receptor related factor 1 neural stem cells gene transfection Parkinson’s disease transplantation rat

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The interaction of Schwann cells with electrospun silk fibroin nanotibers mats

The interaction of Schwann cells with electrospun silk fibro...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Xiao-Li YAN,Ya-Hong ZHAO,Yu-Min YANG~* jiangsu key laboratory of Neuroregeneration,nantong University,nantong,226001,china

＜正＞Objective The bridging of nerve gaps is still one of the major problems in peripheral nerve regeneration.A promising alternative for the repair of peripheral nerve injuries is the bioartificial nerve *** between cultured nerve cells and biomaterials is of *** the purposes of this study,an electrospinning method was used to fabricate silk fibroin（SF） nanofibers mats for cell culture of Schwann Cells（SCs） that were harvested from rats’ sciatic *** Characterizations of alcohol-treated SF electrospun nanofibers were investigated by attenuated total reflection Fourier transform infrared（ATR-FTIR） spectroscopy and scanning electronic microscopy（SEM）.The growth of SCs was observed by light and scanning electron microscopy at regular *** were identified by immunocy-tochemical staining and the viability of SCs was measured by MTT *** SF electrospun nanofibers exhibited a circular cross-section with a smooth *** was a good biological compatibility between SF electrospun nanofibers mats and *** Our results indicated that SF electrospun nanofibers may be a good candidate for the peripheral nerve regeneration applications.

关键词： electrospinning Nanofiber Silk fibroin Schwann cells

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