Evaluation of degree of nerve root injury by dermatomal somatosensory evoked potential following lumbar spinal stenosis

BACKGROUND： Computed Tomography （CT） and Magnetic Resonance Imaging （MRI） can display the site of lumbar spinal stenosis and predict nervous compression at the morphological level; however, pure morphological changes cannot reflect functional alterations in a compressed nerve root. Dermatomal somatosensory evoked potential （DSEP） provides a means to assess the functional state of a nerve root. OBJECTIVE： To evaluate the clinical significance of DSEP, assessing the degree of nerve root injury following lumbar spinal stenosis. DESIGN, TIME AND SETTING： A case-control study was performed in the Department of Orthopaedic Surgery, Hainan People＇s Hospital, China, between September 2004 and December 2007. PARTICIPANTS： Forty-seven patients diagnosed with lumbar spinal stenosis by CT or MRI were selected as the case group; fifty healthy subjects were collected as the control group. METHODS： A KEYPOINT myoelectric evoked potential apparatus （DANTEC Company, Denmark） was used to measure DSEP, and stimulative spots were determined in accordance with the skin key sensory spot standards established by The American Spinal Injury Association： L4 in the medial malleolus, L5 in the third metatarsophalangeal joint of the dorsum of foot and S1 in the lateral heel. The needle electrode used as the recording electrode was located at the Cz point of the cranium, and the reference electrode at the Fz point. MAIN OUTCOME MEASURES： Latency of the P40 peak of DSEP, P1-N1 amplitude, P40 waveform and differentiation and disappearance of various waves. RESULTS： The sensitivity and diagnostic concurrence with surgery of nerve root injury following lumbar spinal stenosis evaluated by DSEP was 95.7 %. P40 latencies at L4, L5 and S1 in the case group were significantly longer than in the control group （P 〈 0.05）, and the P1-N1 amplitude in the case group was significantly lower than the control group （P 〈 0.05-0.01）. Nerve root injury was categorized according to DSEP latency as follows： severe damage （disappearance of the P40 wave in 103 dermatomes）, moderate damage （prolongation of the P40 peak latency ≥ 3.0 times the standard deviation of the normal mean in 60 dermatomes） and mild damage （prolongation of the P40 peak latency ≥ 2.5 times the standard deviation of the normal mean in 31 dermatomes）. CONCLUSION： DSEP can be used to determine the severity of nerve root injury following lumbar spinal stenosis with high sensitivity and specificity.

Reliability of dermatomal somatosensory-evoked potential in the evaluation of lumbosacral nerve root injury A concurrent case-control study

BACKGROUND： It has been reported that dermatomal somatosensory evoked potential （DSEP） can be used for diagnosing nerve root injury in patients with lumbar disc herniation （LDH）, and that 83% 95% of patients suffer from the disease. Body height correction is not performed prior to determinations of latency and latency difference between the healthy and affected sides. However, latency noticeably correlates to body height. OBJECTIVE： This study aims to determine the lumbosacral nerve root injury in patients with LDH by DSEP, and to evaluate the sensitivity of the DSEP difference between the healthy and affected sides using a diagnostic index following body height correction. DESIGN： A case-control observation. SETTING： Department of Orthopedic Surgery, Hainan Provincial People＇s Hospital. PARTICIPANTS： Ninety-six patients, comprised of 67 males and 29 females, with an average age of 43 years and a mean body height of 1.65 m （range 1.48-1.81 m）, were recruited for this study. These patients suffered from unilateral lower limb radiation pain and received treatment at the Department of Orthopedic Surgery, Hainan Provincial People＇s Hospital between January 2004 and December 2006. All patients were confirmed to suffer from LDH at the L3-4, L-5, and/or Ls-SI by CT and/or MRI examinations. Central nervous system diseases were excluded. In order to obtain a normal reference value, DSEP was determined for a group of 50 subjects, who concurrently received health examinations in the same department. The subjects had no previous history of back leg pain or nervous system disease. The group of healthy controls included 26 males and 24 females, with an average age of 37 years and a mean body height of 1.63 m （range 1.50-1.80 m）. Written informed consent was obtained from all subjects for laboratory samples. The protocol was approved by the Hospital＇s Ethics Committees. DSEP was determined with myoelectricity-evoked potential equipment （Keypoint, Batch No. 9020A0042591, Dantec Company, Denmark）. METHODS： DSEP of patients with LDH was determined. Constant-voltage square pulse stimulation was used to determine DSEP, with the following parameters： a pulse wave width of 0.2 milliseconds; a saddle-like stimulating electrode; a stimulation intensity 3 times greater than the sensation threshold; a stimulation frequency of 1.5 Hz; mean superposition greater than 100 times; and inter-electrode impedance 〈 5 k Q. The stimulation point was a skin key sensation point confirmed by the American Spinal Injury Association, i.e. L4 at medial malleolus, L5 at the third metatarsophalangeal joint on the dorsum of the foot, and SI at the lateral heel. The recording electrode was a needle electrode, the recording point was Cz, and the reference electrode was Fz. DSEP latency of P40, and latency differences of P40, between the healthy side and the affected side, were determined. DSEP at L4, L5, and S1 nerve roots of the lower limbs of 50 healthy controls were bilaterally determined. The normal values of P40 latency and P40 N50 amplitude were statistically obtained. MAIN OUTCOME MEASURES： Determination of DSEP values. RESULTS： Ninety-six patients with LDH and fifty healthy controls participated in the final analysis. In the healthy controls, the amplitude of DSEP varied greatly, with a mean amplitude co-efficient of variation of 58% for L4, L5, and SI dermatomes. P40 latency was stable, with a mean latency coefficient of variation of 4.7%. In patients with LDH, the P40 wave disappeared. P40 latency was 2.5 times prolonged compared to normal mean value. P40 latency difference between the healthy and the affected side was 2.5 times higher than the normal mean value of the healthy side. CONCLUSION： DSEP can reflect the functional status of lumbosacral nerve root. P40 latency difference between the healthy side and the affected side is the most sensitive diagnosis index for patients with LDH suffering from unilateral lower limb radiation pain.

Differentiation of endogenous neural stem cells in adult versus neonatal rats after brachial plexus root avulsion injury

An experimental model of brachial plexus root avulsion injury of cervical dorsal C5-6 was established in adult and neonatal rats.Real-time PCR showed that the levels of brain-derived neurotrophic factor,nerve growth factor and neurotrophin-3 in adult rats increased rapidly 1 day after brachial plexus root avulsion injury,and then gradually decreased to normal levels by 21 days.In neonatal rats,levels of the three neurotrophic factors were decreased on the first day after injury,and then gradually increased from the seventh day and remained at high levels for an extended period of time.We observed that greater neural plasticity contributed to better functional recovery in neonatal rats after brachial plexus root avulsion injury compared with adult rats.Moreover, immunohistochemical staining showed that the number of bromodeoxyuridine/nestin-positive cells increased significantly in the spinal cords of the adult rats compared with neonatal rats after brachial plexus root avulsion injury.In addition,the number of bromodeoxyuridine/glial fibrillary acidic protein-positive cells in adult rats was significantly higher than in neonatal rats 14 and 35 days after brachial plexus injury.Bromodeoxyuridine/β-tubulin-positive cells were not found in either adult or neonatal rats.These results indicate that neural stem cells differentiate mainly into astrocytes after brachial plexus root avulsion injury.Furthermore,the degree of neural stem cell differentiation in neonatal rats was lower than in adult rats.

Influence of species,age and weight on the establishment of a murine model of brachial plexus root avulsion

BACKGROUND： Animal models of brachial plexus root avulsion are required for the study of brachial plexus root injuries. The established ventral approach results in slight injuries, and is similar to mechanisms underlying human brachial plexus root avulsion.OBJECTIVE： To analyze the effects of weight, age, and species on the success rate of brachial plexus root avulsion, and to determine the perfect method for establishing models of brachial plexus root avulsion.DESIGN, TIME AND SETTING： A randomized, block design was performed at the Laboratory of Professor Lihua Zhou, Zhongshan School of Medicine, Sun Yat-sen University, China from June 2008 to June 2009.MATERIALS： Sprague Dawley （SD） rats, golden hamsters, and BALb/C mice were used in the present study.METHODS： All animals were randomly subjected to classical brachial plexus root avulsion and modified brachial plexus root avulsion.MAIN OUTCOME MEASURES： Success rate of brachial plexus root avulsion. RESULTS： The success rate of brachial plexus root avulsion was greater in the modified group than in the classical group （P〈0.01）. Moreover, the difference was significant in 15-day-old SD rats, 5-week-old SD rats, and 3-month-old BALb/C mice （P〈0.01）. The success rate of brachial plexus root avulsion was greater in the same weight, 15-day-old juvenile SD rats, than in the 3-month-old BALb/C mice （classical group, P〈0.01; modified group, P〈0.05）. The success rate of brachial plexus root avulsion was significantly greater in 3-month-old golden hamsters than in 5-week-old SD rats in the classical group （P〈0.05）. The success rate of brachial plexus root avulsion was significantly lower in the 15-day-old SD rats compared with the 5-week-old and 3-month-old SD rats in the classical group （P〈0.01）. However, there was no significant difference in the success rate of brachial plexus root avulsion between various ages of SD rats in the modified group （P〉0.05）.CONCLUSION： Modified surgery to induce brachial plexus root avulsion significantly increases the success rate of model establishment. Species, age, and weight affect the success rate of brachial plexus root avulsion, and species plays an important role in the success rate.

The Influence of the Verticillium dahliae Kleb Infection on the Anti-Enzyme Inside the Body of the Cotton with Different Root Injured Degree

This study was to explore the influence of the Verticillium dahliae Kleb inflection on the anti-enzyme inside the body of the cotton with a different root injured degree. When the cotton seedling was long, with four leaves, it was flushed with water carefully, and then the following were obtained： （1） complete root seedling; （2） cut root seedling - by cutting off the lower part, 3-5 cm of the root, with a disinfected knife; （3） injured root seedling - by cutting off most of the side roots, but keeping the main root. Three kinds of cotton seedlings with different roots were immersed separately in different concentrations of the germ liquid （V. dahliae） of 20 mL each. Through 0- 48 h, the wilt degree of the seedling was recorded, and the related anti-enzyme of the variety was measured. After being immersed in the germ liquid, there was a significant difference in the wilt degree of the three kinds of injured root. When the germ liquid was in the ratio of 1：10, the complete root seedling was the lightest with no wilt; the injured root seedling was the second with a 2-degree wilt; but the cut root seedling was the most serious with a 3- degree wilt. At the same time, the changes in the peroxidase and malondialdehvde activities were determined. Peroxidase （POD） activities in the cut root seedling were 38.2 U mg^-1 min^-1, in the injured root seedling were 42.96 U mg^-1 min^-1, and in the complete root seedling were the highest at 49.2 U mg^-1 min^-1. The malondialdenvde （MDA） content in cut root seedling was 39.483 mmol g^-1, injured root seedling was 27.12 mmol g^-1, and the complete root seedling was only 3.845 mmol g^-1 The activity of the related anti-enzymes, such as POD was high or low, the quantity of the MDA was more or less, which they met the order of the harm of the seedlings. The change of SOD activities in cut root seedling was the most obvious as well. After injuring and inflecting the young roots, the exterior pathological reaction of the seedling and the dynamic state biochemical reaction of the related enzymes inside the plant body were studied. It showed that the plant exterior pathology responded to the test, with the internal biochemical reaction fitting together mutually.

Puerarin ameliorates allodynia and hyperalgesia in rats with peripheral nerve injury

Puerarin is a major active ingredient of the traditional Chinese plant medicine,Radix Puerariae,and commonly used in the treatment of myocardial and cerebral ischemia.However,the effects of puerarin on neuropathic pain are still unclear.In this study,a neuropathic pain animal model was created by partial sciatic nerve ligation.Puerarin（30 or 60 mg/kg） was intraperitoneally injected once a day for 7 days.Mechanical allodynia and thermal hyperalgesia were examined at 1 day after model establishment.Mechanical threshold and paw withdrawal latency markedly increased in a dose-dependent manner in puerarin-treated rats,especially at 7 days after model establishment.At 7 days after model establishment,quantitative real-time reverse transcriptase-polymerase chain reaction results showed that puerarin administration reversed m RNA expression of transient receptor potential vanilloid 1（Trpv1） and transient receptor potential ankyrin 1（Trpa1） in a dose-dependent manner in dorsal root ganglion neurons after peripheral nerve injury.These results suggest that puerarin dose-dependently ameliorates neuropathic pain by suppressing Trpv1 and Trpa1 up-regulation in dorsal root ganglion of neuropathic pain rats.

Effects of Root Cutting on Morphological Characteristics and Endogenous Hormone Levels of Quercus variabilis Seedlings

The objective of this study was to investigate the effects of root cutting stress on the dynamic changes of endogenous hormone content and growth characteristics of Quercus variabilis roots,and to explore the physiological role of endogenous hormones in regulating root-crown interactions in the short term.The morphological characteristics and endogenous hormone contents of normal roots(no root cutting,CK)and cut roots(cut by 1/3 of the length of the main root,RP)were determined by liquid chromatography,which was combined with mass spectrometry at different levels of different developmental stages.The results showed that the root growth indexes and root endogenous hormones in the RP group were superior to those in the CK group.Through comprehensive analysis of endogenous hormones,it was found that the crosstalk of IAA,JA,ABA and SA could activate the root growth defense.After the root cutting treatment,the root growth of Quercus variabilis seedlings could compensate for the inhibition of taproot growth by promoting lateral root growth.The growth and development of compensatory lateral roots contribute to the increase the total root length of plants,thus promoting the absorption of water and nutrients.It is speculated that plant hormones may be the key factors affecting their development,but this is not only related to the content of a single hormone,but more importantly,it is the interaction between various hormones.

Microencapsulation improves inhibitory effects of transplanted olfactory ensheathing cells on pain after sciatic nerve injury

Olfactory bulb tissue transplantation inhibits P2X2/3 receptor-mediated neuropathic pain. However, the olfactory bulb has a complex cellular composition, and the mechanism underlying the action of purified transplanted olfactory ensheathing cells（OECs） remains unclear. In the present study, we microencapsulated OECs in alginic acid, and transplanted free and microencapsulated OECs into the region surrounding the injured sciatic nerve in rat models of chronic constriction injury. We assessed mechanical nociception in the rat models 7 and 14 days after surgery by measuring paw withdrawal threshold, and examined P2X2/3 receptor expression in L4–5 dorsal root ganglia using immunohistochemistry. Rats that received free and microencapsulated OEC transplants showed greater withdrawal thresholds than untreated model rats, and weaker P2X2/3 receptor immunoreactivity in dorsal root ganglia. At 14 days, paw withdrawal threshold was much higher in the microencapsulated OEC-treated animals. Our results confirm that microencapsulated OEC transplantation suppresses P2X2/3 receptor expression in L4–5 dorsal root ganglia in rat models of neuropathic pain and reduces allodynia, and also suggest that transplantation of microencapsulated OECs is more effective than transplantation of free OECs for the treatment of neuropathic pain.

Implanting iodine-125 seeds into rat dorsal root ganglion for neuropathic pain: neuronal microdamage without impacting hind limb motion

The use of iodine-125 （L251） in cancer treatment has been shown to relieve patients＇ pain. Consid- ering dorsal root ganglia are critical for neural transmission between the peripheral and central nervous systems, we assumed that 125I could be implanted into rat dorsal root ganglia to provide relief for neuropathic pain. 125I seeds with different radioactivity （0, 14.8, 29.6 MBq） were im- planted separately through L4-5 and L5-6 intervertebral foramen into the vicinity of the L5 dorsal root ganglion, von Frey hair results demonstrated the mechanical pain threshold was elevated after implanting 125I seeds from the high radioactivity group. Transmission electron microscopy revealed that nuclear membrane shrinkage, nucleolar margination, widespread mitochondrial swelling, partial vacuolization, lysosome increase, and partial endoplasmic reticulum dilation were visible at 1,440 hours in the low radioactivity group and at 336 hours in the high radio- activity group. Abundant nuclear membrane shrinkage, partial fuzzy nuclear membrane and endoplasmic reticulum necrosis were observed at 1,440 hours in the high radioactivity group. No significant difference in combined behavioral scores was detected between preoperation and postoperation in the low and high radioactivity groups. These results suggested that the mechan- ical pain threshold was elevated after implanting 125I seeds without influencing motor functions of the hind limb, although cell injury was present.

Treatment with analgesics after mouse sciatic nerve injury does not alter expression of wound healingassociated genes

Animal models of sciatic nerve injury are commonly used to study neuropathic pain as well as axon regeneration. Administration of post-surgical analgesics is an important consideration for animal welfare, but the actions of the analgesic must not interfere with the scientific goals of the experiment. In this study, we show that treatment with either buprenorphine or acetaminophen following a bilateral sciatic nerve crush surgery does not alter the expression in dorsal root ganglion（DRG） sensory neurons of a panel of genes associated with wound healing. These findings indicate that the post-operative use of buprenorphine or acetaminophen at doses commonly suggested by Institutional Animal Care and Use Committees does not change the intrinsic gene expression response of DRG neurons to a sciatic nerve crush injury, for many wound healing-associated genes. Therefore, administration of post-operative analgesics may not confound the results of transcriptomic studies employing this injury model.

End-to-side neurorrhaphy repairs peripheral nerve injury：sensory nerve induces motor nerve regeneration

End-to-side neurorrhaphy is an option in the treatment of the long segment defects of a nerve.It involves suturing the distal stump of the disconnected nerve（recipient nerve） to the side of the intimate adjacent nerve（donor nerve）.However,the motor-sensory specificity after end-to-side neurorrhaphy remains unclear.This study sought to evaluate whether cutaneous sensory nerve regeneration induces motor nerves after end-to-side neurorrhaphy.Thirty rats were randomized into three groups：（1） end-to-side neurorrhaphy using the ulnar nerve（mixed sensory and motor） as the donor nerve and the cutaneous antebrachii medialis nerve as the recipient nerve;（2） the sham group：ulnar nerve and cutaneous antebrachii medialis nerve were just exposed;and（3） the transected nerve group：cutaneous antebrachii medialis nerve was transected and the stumps were turned over and tied.At 5 months,acetylcholinesterase staining results showed that 34% ± 16% of the myelinated axons were stained in the end-to-side group,and none of the myelinated axons were stained in either the sham or transected nerve groups.Retrograde fluorescent tracing of spinal motor neurons and dorsal root ganglion showed the proportion of motor neurons from the cutaneous antebrachii medialis nerve of the end-to-side group was 21% ± 5%.In contrast,no motor neurons from the cutaneous antebrachii medialis nerve of the sham group and transected nerve group were found in the spinal cord segment.These results confirmed that motor neuron regeneration occurred after cutaneous nerve end-to-side neurorrhaphy.

Central projections and connections of lumbar primary afferent fibers in adult rats:effectively revealed using Texas red-dextran amine tracing

Signals from lumbar primary afferent fibers are important for modulating locomotion of the hind-limbs.However,silver impregnation techniques,autoradiography,wheat germ agglutinin-horseradish peroxidase and cholera toxin B subunit-horseradish peroxidase cannot image the central projections and connections of the dorsal root in detail.Thus,we injected 3-k Da Texas red-dextran amine into the proximal trunks of L4 dorsal roots in adult rats.Confocal microscopy results revealed that numerous labeled arborizations and varicosities extended to the dorsal horn from T12–S4,to Clarke＇s column from T10–L2,and to the ventral horn from L1–5.The labeled varicosities at the L4 cord level were very dense,particularly in laminae I–Ⅲ,and the density decreased gradually in more rostral and caudal segments.In addition,they were predominately distributed in laminae I–IV,moderately in laminae V–VⅡ and sparsely in laminae VⅢ–X.Furthermore,direct contacts of lumbar afferent fibers with propriospinal neurons were widespread in gray matter.In conclusion,the projection and connection patterns of L4 afferents were illustrated in detail by Texas red-dextran amine-dorsal root tracing.