Protective effect of thodioloside and bone marrow mesenchymal stem cells infected with HIF-1-expressing adenovirus on acute spinal cord injury

Rhodioloside has been shown to protect cells from hypoxia injury,and bone marrow mesenchymal stem cells have a good effect on tissue repair.To study the effects of rhodioloside and bone marrow mesenchymal stem cells on spinal cord injury,a rat model of spinal cord injury was established using the Infinite Horizons method.After establishing the model,the rats were randomly divided into five groups.Rats in the control group were intragastrically injected with phosphate buffered saline(PBS)(5μL).PBS was injected at 6 equidistant points around 5 mm from the injury site and at a depth of 5 mm.Rats in the rhodioloside group were intragastrically injected with rhodioloside(5 g/kg)and intramuscularly injected with PBS.Rats in the mesenchymal stem cell(MSC)group were intramuscularly injected with PBS and intramuscularly with MSCs(8×10^6/mL in a 50-μL cell suspension).Rats in the Ad-HIF-MSC group were intragastrically injected with PBS and intramuscularly injected with HIF-1 adenovirus-infected MSCs.Rats in the rhodioloside+Ad-HIF-MSC group were intramuscularly injected with MSCs infected with the HIF-1 adenovirus and intragastrically injected with rhodioloside.One week after treatment,exercise recovery was evaluated with a modified combined behavioral score scale.Hematoxylin-eosin staining and Pischingert’s methylene blue staining were used to detect any histological or pathological changes in spinal cord tissue.Levels of adenovirus IX and Sry mRNA were detected by real-time quantitative polymerase chain reaction and used to determine the number of adenovirus and mesenchymal stem cells that were transfected into the spinal cord.Immunohistochemical staining was applied to detect HIF-1 protein levels in the spinal cord.The results showed that:(1)compared with the other groups,the rhodioloside+Ad-HIF-MSC group exhibited the highest combined behavioral score(P<0.05),the most recovered tissue,and the greatest number of neurons,as indicated by Pischingert’s methylene blue staining.(2)Compared with the PBS group,HIF-1 protein expression was greater in the rhodioloside group(P<0.05).(3)Compared with the Ad-HIF-MSC group,Sry mRNA levels were higher in the rhodioloside+Ad-HIF-MSC group(P<0.05).These results confirm that rhodioloside combined with bone marrow mesenchymal stem cells can promote the recovery of spinal cord injury and activate the HIF-1 pathway to promote the survival of bone marrow mesenchymal stem cells and repair damaged neurons within spinal cord tissue.This experiment was approved by the Animal Ethics Committee of Gansu University of Traditional Chinese Medicine,China(approval No.2015KYLL029)in June 2015.

Factors related to improved American Spinal Injury Association grade of acute traumatic spinal cord injury

BACKGROUND Acute traumatic spinal cord injury(ATSCI)usually results in disability,yet data on contemporary national trends of ATSCI incidence are limited.AIM To provide a systematic and basic theoretical basis for improving the treatment of acute spinal cord injury.METHODS Data from the Peking University Third Hospital Inpatient Sample databases were analyzed.A total of 304 patients with ATSCI were included from 2012 to 2017.The epidemiological data,treatment,complications and clinical outcomes of these patients were reviewed.RESULTS Of the 304 patients,257(84.5%)were male,and 75%of the patients were 55 years old or younger.135 patients had improved follow-up American Spinal Injury Association(ASIA)grades(44.4%).Only 14 patients with ASIA grade A improved.A statistically significant difference in prognosis between patients who underwent surgery within 72 h and those who underwent surgery after 72 h was observed(P<0.05).Surgery within 72 h resulted in better prognosis.The Steroid group and the Non-Steroid group showed a significant difference in outcome among patients with ASIA grades A and B(P<0.05).Patients with pneumonia had a poorer prognosis than patients without pneumonia(P<0.05).Surgery within 72 h resulted in better prognosis.CONCLUSION This study found that there was no significant difference in hospitalization time and prognosis between the Steroid group and the Non-Steroid group,but the patients with severe spinal cord injury(ASIA grades A and B)who underwent surgery combined with steroid therapy had a better prognosis than those who underwent surgery alone.The disastrous consequences of ATSCI and lack of consensus on the management strategy are obvious.Further improvements in treatment planns are needed in order to obtain more reliable functional outcomes.

Effects of decompression joint Governor Vessel electro-acupuncture on rats with acute upper cervical spinal cord injury

Decompression is the major therapeutic strategy for acute spinal cord injury,but there is some debate about the time window for decompression following spinal cord injury.An important goal and challenge in the treatment of spinal cord injury is inhibiting or reversing secondary injury.Governor Vessel electroacupuncture can improve symptoms of spinal cord injury by inhibiting cell apoptosis and improving the microenvironment of the injured spinal cord.In this study,Governor Vessel electroacupuncture combined with decompression at different time points was used to treat acute spinal cord injury.The rat models were established by inserting a balloon catheter into the atlanto-occipital space.The upper cervical spinal cord was compressed for 12 or 48 hours prior to decompression.Electroacupuncture was conducted at the acupoints Dazhui（GV14） and Baihui（GV 20）（2 Hz,15 minutes） once a day for 14 consecutive days.Compared with decompression alone,hind limb motor function recovery was superior after decompression for 12 and 48 hours combined with electroacupuncture.However,the recovery of motor function was not significantly different at 14 days after treatment in rats receiving decompression for 12 hours.Platelet-activating factor levels and caspase-9 protein expression were significantly reduced in rats receiving electroacupuncture compared with decompression alone.These findings indicate that compared with decompression alone,Governor Vessel electroacupuncture combined with delayed decompression（48 hours） is more effective in the treatment of upper cervical spinal cord injury.Governor Vessel electroacupuncture combined with early decompression（12 hours） can accelerate the recovery of nerve movement in rats with upper cervical spinal cord injury.Nevertheless,further studies are necessary to confirm whether it is possible to obtain additional benefit compared with early decompression alone.

Management of acute spinal cord injury:A summary of the evidence pertaining to the acute management,operative and non-operative management

Acute traumatic spinal cord injury is often a lifechanging and devastating event with considerable mortality and morbidity.Over half a million people suffer from traumatic spinal cord injury annually with the majority resulting from road traffic accidents or falls.The Individual,societal and economic costs are enormous.Initial recognition and treatment of acute traumatic spinal cord injury are crucial to limit secondary injury to the spinal cord and to provide patients with the best chance of some functional recovery.This article is an overview of the management of the acute traumatic spinal cord injury patient presenting to the emergency department.We review the initial assessment,criteria for imaging and clearing the spine,and evaluate the literature to determine the optimum timing of surgery and the role of non-surgical treatment in patients presenting with acute spinal cord injury.

Future directions for using estrogen receptor agonists in the treatment of acute and chronic spinal cord injury

All synthetic and natural estrogen receptor agonists, in- cluding the most potent physiological molecule estrogen or estradiol （E2）, work typically via activation of nuclear estrogen receptor alpha （ERα） and estrogen receptor beta （ERβ）. Both ERα and ERβ modulate the expression of a variety of genes in the cells. Neurons and glial cells express ERa and ERβ. Many studies so far from our and other laboratories have firmly established the mode of actions that ERα and ERβ agonists are very promising anti-inflammatory and neuroprotective agents in the treatment of neurodegenera- rive diseases and injuries including spinal cord injury （SCI） （Chakrabarti et al., 2014a）.

Early methylprednisolone impact treatment for sensory and motor function recovery in patients with acute spinal cord injury A self-control study

BACKGROUND： For the treatment of spinal cord injury, any pathological changes of the injured tissue should be primarily corrected or reversed. Any remaining fibrous function and neurons with intact structure should be retained, and the toxic substances caused by ischemia-hypoxia following spinal cord injury, should be eliminated to create a favorable environment that would promote neural functional recovery. OBJECTIVE： This study was designed to investigate the effects of the impact of early methylprednisolone-treatment on the sensory and motor function recovery in patients with acute spinal cord injury. DESIGN： A self-control observation. SETTING： Department of Spine Surgery, First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China. PARTICIPANTS： Forty-three patients with acute spinal cord injury were admitted to the Department of Spine Surgery, First Affiliated Hospital of Nanjing Medical University, between October 2005 and September 2007. These patients were recruited for the present study. The patients comprised 33 males and 10 females, and all met with the inclusive criteria namely, the time between suffering from acute spinal cord injury and receiving treatment was less than or equal to eight hours. METHODS： According to the protocol determined by the State Second Conference of Acute Spinal Cord Injury of USA, all patients received the drop-wise administration of a 30-mg/kg dose of methylprednisolone （H200040339, 500 mg/bottle, Pharmacia N.V/S.A, Belgium） for 15 minutes within 8 hours post injury. After a 45-minute interval, methylprednisolone was administered at 5.4 mg/kg/h for 23 hours. MAIN OUTCOME MEASURES： Prior to and post treatment, acupuncture sense and light touch scoring were performed at 28 dermatomic area key points, including occipital tuberosity and supraclavicular fossa. At the same time, motor scoring of key muscles among 10 pairs of sarcomeres was also performed. RESULTS： All 43 patients participated in the final analysis. There was no significant difference of sensory and motor scores in patients with complete acute spinal cord injury between prior to and post methylprednisolone impact treatment （P 〉 0.05）. The motor score was significantly decreased in patients with incomplete acute spinal cord injury post methylprednisolone impact treatment （P 〈 0.01 ）. CONCLUSION： Early methylprednisolone impact may improve the motor function of patients with incomplete acute spinal cord injury. However, it has no influences on patients with complete acute spinal cord injury.

Provision of nutrients after acute spinal cord injury: the implications of feast and famine

Traumatic spinal cord injury（SCI）often leaves patients with devastating neurological deficits.The traumatic event–or primary injury–can be due to mechanisms such as compression,distraction,shear,laceration or（rarely）even transection.Thereafter SCI patients are vulnerable to progressive,delayed damage as a result of secondary insults and secondary injury.Secondary insults such as hypoxia and hypotension occur at the level of the organism from a myriad of causes.

Effect of electroacupuncture at Jiaji (EX-B2) points on the expression of microRNA-21 and neuronal apoptosis in rats with acute spinal cord injury

Objective:To observe the effect of electroacupuncture at Jiaji (EX-B2) points on the expression of microRNA-21 and neuronal apoptosis in rats with acute spinal cord injury. Methods:SD rats were randomly divided into sham-operated group, model group, electro-acupuncture group and methylprednisolone group. The model of acute spinal cord injury was reproduced by modified Allen's method. Each group was given corresponding intervention treatment 2 hours after modeling. The electroacupuncture group was treated with electro-acupuncture at Jiaji (EX-B2) points in T8 to T12, the methylprednisolone group was given intraperitoneal injection of 30 mg/kg methylprednisolone sodium succinate, and the sham operation group and model group were not treated. After treatment, BBB scores were observed to evaluate the motor function of posterior limb in each group, and the injured spinal cord tissues were taken. The pathological changes of spinal cord neuropathy were observed by Nissl staining. The expression of microRNA-21 was detected by RT-qPCR, and the expression of apoptotic proteins (Bax, Bcl-2 and cleaved-Caspase-3) was detected by Western-Blot. Result:Compared with sham operation group, BBB, neuron survival rate and expression of microRNA-21 were significantly lower, apoptotic rate of neurons was significantly higher, expression of Bax, Bcl-2, cleaved Caspase-3 protein and Bax/Bcl-2 ratio were significantly increased in model group (P< 0.05). Electroacupuncture could significantly increase BBB score, neuron survival rate and expression of microRNA-21, reduce apoptotic rate of neurons and the ratio of Bax/Bcl-2, and inhibit the expression of Bax, Bcl-2 and cleaved Caspase-3 protein in tissues, which were significantly different from those of model group (P< 0.05). Conclusions: Electroacupuncture at Jiaji (EX-B2) points can significantly promote the recovery of neurological function in rats after ASCI, and its mechanism may be closely related to the up-regulation of the expression of microRNA-21 in tissues and the inhibition of the activation of Bax/Bcl-2/cleaved Caspase-3 signaling pathway.

Effect of FTY720 on RhoA expression after acute spinal cord injury in rats

Objective To investigate the effect of fingolimod (FTY720)on RhoA expression after spinal cord injury (SCI)in rats,and explore the possible mechanism of FTY720in the treatment of SCI.Methods A rat model of acute SCI was established with a

Effects of low dose Glibenclamide on secondary damage after acute spinal cord injury in rats

Objective To investigate the effects of Glibenclamide on reduction of secondary damage after acute spinal cord injury in rats.Methods Ninety rats were randomly divided into control group(laminectomy alone),spinal cord injury group(injury group),and treatment group(treated

Neuroprotective effects of rapamycin on spinal cord injury in rats by increasing autophagy and Akt signaling

Rapamycin treatment has been shown to increase autophagy activity and activate Akt phosphorylation, suppressing apoptosis in several models of ischemia reperfusion injury. However, little has been studied on the neuroprotective effects on spinal cord injury by activating Akt phosphorylation. We hypothesized that both effects of rapamycin, the increased autophagy activity and Akt signaling, would contribute to its neuroprotective properties. In this study, a compressive spinal cord injury model of rat was created by an aneurysm clip with a 30 g closing force. Rat models were intraperitoneally injected with rapamycin 1 mg/kg, followed by autophagy inhibitor 3-methyladenine 2.5 mg/kg and Akt inhibitor IV 1 μg/kg. Western blot assay, immunofluorescence staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay were used to observe the expression of neuronal autophagy molecule Beclin 1, apoptosis-related molecules Bcl-2, Bax, cytochrome c, casp ase-3 and Akt signaling. Our results demonstrated that rapamycin inhibited the expression of mTOR in injured spinal cord tissue and up-regulated the expression of Beclin 1 and phosphorylated-Akt. Rapamycin prevented the decrease of bcl-2 expression in injured spinal cord tissue, reduced Bax, cytochrome c and caspase-3 expression levels and reduced the number of apoptotic neurons in injured spinal cord tissue 24 hours after spinal cord injury. 3-Methyladenine and Akt inhibitor IV intervention suppressed the expression of Beclin-1 and phosphorylated-Akt in injured spinal cord tissue and reduced the protective effect of rapamycin on apoptotic neurons. The above results indicate that the neuroprotective effect of rapamycin on spinal cord injury rats can be achieved by activating autophagy and the Akt signaling pathway.

Current therapeutic strategies for inflammation following traumatic spinal cord injury

Damage from spinal cord injury occurs in two phases-the trauma of the initial mechanical insult and a secondary injury to nervous tissue spared by the primary insult.Apart from damage sustained as a result of direct trauma to the spinal cord,the post-traumatic inflammatory response contributes significantly to functional motor deficits exacerbated by the secondary injury.Attenuating the detrimental aspects of the inflammatory response is a promising strategy to potentially ameliorate the secondary injury,and promote significant functional recovery.This review details how the inflammatory component of secondary injury to the spinal cord can be treated currently and in the foreseeable future.

Identification of potential oxidative stress biomarkers for spinal cord injury in erythrocytes using mass spectrometry

Oxidative stress is a hallmark of secondary injury associated with spinal cord injury.Identifying stable and specific oxidative biomarkers is of important significance for studying spinal cord injury-associated secondary injury.Mature erythrocytes do not contain nuclei and mitochondria and cannot be transcribed and translated.Therefore, mature erythrocytes are highly sensitive to oxidative stress and may become a valuable biomarker.In the present study, we revealed the proteome dynamics of protein expression in erythrocytes of beagle dogs in the acute and subacute phases of spinal cord injury using mass spectrometry-based approaches.We found 26 proteins that were differentially expressed in the acute(0–3 days) and subacute(7–21 days) phases of spinal cord injury.Bioinformatics analysis revealed that these differentially expressed proteins were involved in glutathione metabolism, lipid metabolism, and pentose phosphate and other oxidative stress pathways.Western blot assays validated the differential expression of glutathione synthetase, transaldolase, and myeloperoxidase.This result was consistent with mass spectrometry results, suggesting that erythrocytes can be used as a novel sample source of biological markers of oxidative stress in spinal cord injury.Glutathione synthetase, transaldolase, and myeloperoxidase sourced from erythrocytes are potential biomarkers of oxidative stress after spinal cord injury.This study was approved by the Experimental Animal Centre of Ningxia Medical University, China(approval No.2017-073) on February 13, 2017.

Surgical decompression in acute spinal cord injury： A review of clinical evidence, animal model studies, and potential future directions of investigation

The goal for treatment in acute spinal cord injury （SCI） is to reduce the extent of secondary damage and facilitate neurologic regeneration and functional recovery. Although multiple studies have investigated potential new therapies for the treatment of acute SCI, outcomes and management protocols aimed at ameliorating neurologic injury in patients remain ineffective. More recent clinical and basic science research have shown surgical interventions to be a potentially valuable modality for treatment; however, the role and timing of surgical decompression, in addition to the optimal surgical intervention, remain one of the most controversial topics pertaining to surgical treatment of acute SCI. As an increasing number of potential treatment modalities emerge, animal models are pivotal for investigating its clinical application and translation into human trials. This review critically appraises the available literature for both clinical and basic science studies to highlight the extent of investigation that has occurred, specific therapies considered, and potential areas for future research.

Pros and Cons: Autophagy in Acute Spinal Cord Injury

Autophagy plays an important role in the development and pathogenesis of various diseases. It can be induced by a variety of events such as hypoxia, nutrient-starvation, and mechanical damage. Many neurological disorders such Parkinson’s disease, Alzheimer’s disease, amyotrophic lateral sclerosis, Huntington’s disease, cerebral ischemia, and acute spinal cord injury (ASCI), are closely related to autophagy. However, therapeutic strategies to manipulate autophagy have not yet been fully deciphered due to the limited knowledge of the molecular mechanisms underlying autophagy in these disorders.

Guidelines for management of pediatric acute hyperextension spinal cord injury

Pediatric acute hyperextension spinal cord injury(SCI)named as PAHSCI by us,is a special type of thoracolumbar SCI without radiographic abnormality and highly related to back-bend in dance training,which has been increasingly reported.At present,it has become the leading cause of SCI in children,and brings a heavy social and economic burden.Both domestic and foreign academic institutions and dance education organizations lack a correct understanding of PAHSCI and relevant standards,specifications or guidelines.In order to provide standardized guidance,the expert team formulated this guideline based on the principles of science and practicability,starting from the diagnosis,differential diagnosis,etiology,admission evaluation,treatment,complications and prevention.This guideline puts forward 23 recommendations for 14 related issues.

夹脊电针联合甲基强的松龙对ASCI大鼠脊髓组织NMDA受体亚单位NR1蛋白表达的影响

目的:观察夹脊电针联合甲基强的松龙对急性脊髓损伤大鼠肢体运动功能评分及脊髓组织中N-甲基-D-天冬氨酸受体亚单位NR1蛋白表达的影响,探讨夹脊电针联合甲基强的松龙治疗ASCI的作用机制。方法:将72只雌性Wistar大鼠随机分为假手术组(Sham)、模型组(ASCI)和夹脊电针联合甲基强的松龙组(EA+MP),又分为1天、3天、7天和14天4个亚组,每个亚组6只。NYU打击器制备急性脊髓损伤大鼠模型。Sham组进行手术,不造模,术后常规饲养;ASCI组造模后常规饲养;EA+MP组造模后30 min,给予大鼠一次性注射甲基强的松龙30 mg/kg,术后3 h给予大鼠夹脊电针治疗,每次30 min,每日1次。采用BBB评分观察ASCI大鼠肢体运动功能,免疫组化法检测各组大鼠脊髓组织NMDA-NR1蛋白表达。结果:与Sham组比较,ASCI组大鼠BBB评分显著降低(P<0.05);与ASCI组比较,经夹脊电针联合甲基强的松龙治疗3天后,EA+MP组BBB评分显著升高(P<0.05)。与Sham组比较,ASCI组大鼠脊髓组织NMDA-NR1平均光密度值升高(P<0.05);治疗3天后,与ASCI组相比,EA+MP组大鼠脊髓组织NMDA-NR1平均光密度值降低(P<0.05)。结论:急性脊髓损伤后大鼠脊髓组织NMDA-NR1明显增高,夹脊电针联合甲基强的松龙治疗后NMDA-NR1显著下调,可能是其改善ASCI大鼠肢体运动功能的机制之一。

夹脊电针联合甲基强的松龙(MP)对ASCI大鼠脊髓组织病理形态学影响的时效关系实验研究

目的探寻夹脊电针联合甲基强的松龙(MP)治疗对大鼠急性脊髓损伤(ASCI)后组织的病理学改变影响的时效关系。方法将72只大鼠雌性Wistar大鼠按随机数字表法平均分为Sham组、ASCI组、EA+MP组,每组又分为1 d、3 d、7 d、14 d 4个亚组。采用BBB评分观察不同治疗方法在不同时间点对ASCI大鼠肢体运动功能的影响;采用HE染色观察在各个时间点对ASCI大鼠脊髓组织病理学影响;比较Sham组、ASCI组、EA+MP组在不同的时间点治疗效果,筛选最佳治疗方案。结果 BBB评分结果显示,术后3 d、7 d、14 d,与ASCI组比较,EA+MP组大鼠肢体运动功能评分升高(P<0.05),EA+MP组组间比较,14 d组明显优于3 d组;HE染色结果显示ASCI组大鼠1 d时出血明显,可见大量凋亡神经细胞;3 d时细胞破坏最明显,7 d时神经细胞凋亡有所减轻,神经细胞坏死,4 d时凋亡已不明显,可见空泡存在,治疗组神经细胞损伤情况较ASCI组明显减轻,神经细胞保存较完好。结论夹脊电针联合MP能改善ASCI大鼠运动功能评分,同时减少细胞凋亡,减轻脊髓病理学形态改变,并随着时间延长效果愈加明显。

促红细胞生成素对大鼠急性脊髓损伤铜死亡及感觉运动功能的影响

目的探讨促红细胞生成素(EPO)对大鼠急性脊髓损伤(ASCI)铜死亡及感觉运动功能的影响。方法将36只212~236 g雄性SD大鼠按照随机数字表法分为Sham组(行椎板切除)、ASCI组(ASCI建模)和EPO组(行ASCI建模后每日腹腔注射EPO 5000 U/kg,7 d),每组12只。术后进行运动及感觉功能(Reuter)评分、BBB评分及脊髓含水率测量,采用等离子质谱仪检测Cu^(2+)含量,蛋白质印迹技术检测铜死亡蛋白铁氧还原蛋白1(FDX1)及二氢硫辛酰转乙酰基酶(DLAT)的表达水平,比较3组大鼠感觉运动功能、脊髓水肿情况等。结果与Sham组比较,ASCI组和EPO组BBB评分减少(P<0.05),Reuter评分、脊髓含水率、Cu^(2+)含量、FDX1及DLAT表达均增多(P<0.05);与ASCI组比较,EPO组大鼠BBB评分增多(P<0.05),Reuter评分、脊髓含水率、Cu^(2+)含量、FDX1及DLAT表达均减少(P<0.05)。结论EPO可抑制ASCI大鼠铜死亡、减轻脊髓水肿,改善其感觉运动功能。

鼠神经生长因子(mNGF)联合康复训练治疗急性脊髓损伤(ASCI)患者的临床效果分析

目的探讨分析为急性脊髓损伤患者实施鼠神经生长因子药物治疗联合康复训练治疗的临床效果.方法2019年1月—2020年2月,将该院60例急性脊髓损伤患者随机等分为两组,每组30例,为参照组行单纯性鼠神经生长因子药物治疗,针对研究组实施鼠神经生长因子药物治疗联合康复训练治疗,对比两组的治疗有效率以及接受治疗前后的ASIA评分指标.结果研究组的治疗有效率高于参照组(P<0.05).治疗前,研究组的运动评分指标、针刺评分指标以及触觉评分指标均与参照组差异无统计学意义(P>0.05).治疗后,研究组的运动评分指标、针刺评分指标以及触觉评分指标均高于参照组(P<0.05).结论为急性脊髓损伤患者实施鼠神经生长因子药物治疗联合康复训练治疗,能获取较好效果,值得临床推广.