Neurological effects of recombinant human erythropoietin in Friedreich's ataxia: A clinical pilot trial

In a “proof‐of‐concept” study, we demonstrated that recombinant human erythropoietin (rhuEPO) increases frataxin levels in Friedreich's ataxia (FRDA) patients. We now report a 6‐month open‐label clinical pilot study of safety and efficacy of rhuEPO treatment in FRDA. Eight adult FRDA patients received 2.000 IU rhuEPO thrice a week subcutaneously. Clinical outcome measures included Ataxia Rating Scales. Frataxin levels and indicators for oxidative stress were assessed. Hematological parameters were monitored biweekly. Scores in Ataxia Rating Scales such as FARS (P = 0.0063) and SARA (P = 0.0045) improved significantly. Frataxin levels increased (P = 0.017) while indicators of oxidative stress such as urine 8‐OHdG (P = 0.012) and peroxide levels decreased (P = 0.028). Increases in hematocrit requiring phlebotomies occurred in 4 of 8 patients. In this explorative open‐label clinical pilot study, we found an evidence for clinical improvement together with a persistent increase of frataxin levels and a reduction of oxidative stress parameters in patients with FRDA receiving chronic treatment with rhuEPO. Safety monitoring with regular blood cell counts and parameters of iron metabolism is a potential limitation of this approach. © 2008 Movement Disorder Society     

Skeletal muscle involvement in human immunodeficiency virus infection

Summary In addition to muscle changes due to peripheral nervous system involvement, primary myopathic changes associated with the human immunodeficiency virus (HIV) have also been described. We studied seven cases: two had developed an acquired immunodeficiency syndrome (AIDS) four had seroconverted to HIV but were otherwise asymptomatic, one was HIV seronegative when the biopsy was performed and one was biopsied twice. Besides the HIV no other infectious agent was detected. Muscle biopsies showed: (a) muscle fiber necrosis and regeneration; (b) inflammatory changes with moderate perivascular infiltration; and (c) unusual myofibrillary disorganization. Immunocytochemical techniques using anti-HIV monoclonal antibodies showed the presence of the virus in one biopsy. HIV-RNA was detected by in situ hybridization in the same biopsy. With both techniques the HIV was detected in isolated mononuclear cells in the muscle endomysium and not within the muscle fibers. Muscle involvement associated with HIV infection may be related, at least in some cases, to the presence of the virus in interstitial cells.Supported by a grant from Agence Nationale de Recherches sur le SIDA (A.N.R.S.)Presented in part at the joint MRC/INSERM meeting on HIV infection and the nervous system, Paris, France, January 19–20, 1989     

Haemostatic effects of recombinant human erythropoietin in chronic haemodialysis patients

Recombinant human erythropoietin was administered for up to 40 weeks to nine patients on chronic haemodialysis. From the third week of administration onwards, not only haemoglobin and haematocrit but also the platelet count rose, the latter, however, transiently. Subnormal platelet aggregation before therapy also improved transiently and in parallel with the erythropoietin dosage. The bleeding time normalized in almost all patients. There were no major side-effects. We conclude that recombinant erythropoietin improves haemostasis in chronic haemodialysis patients by increasing the haematocrit and in addition transiently enhances platelet number and function.     

Skeletal muscle involvement in human immunodeficiency virus infection: a report of four cases

Human immunodeficiency virus (HIV) may affect any part of the neuraxis and may affect skeletal muscle in many ways, ranging from myofiber atrophy in the wasting syndrome to inflammatory muscle disease and a host of opportunistic infections involving muscle. We report here a case series of 4 zidovudine-naοve patients with proven HIV infection with myopathy. One was a case of HIV wasting syndrome, and the three others were diagnosed as HIV polymyositis. Muscle biopsy proved invaluable in the characterization of these cases.     

Neuroprotective effects of recombinant human erythropoietin on facial motoneurons after facial nerve injury in rats★

BACKGROUND： Erythropoietin and recombinant human erythropoietin （rhEPO） inhibit apoptosis of motor neurons caused by spinal cord injury and brain damage in rats. However, it still remains to be shown whether rhEPO can protect facial motoneurons （FMNs） as Well. OBJECTIVE： To test the neuroprotective effects of rhEPO on injured VMNs, as well as the influence on Caspase-3 expression. DESIGN, TIME AND SETTING： Randomized, controlled, animal experiment. This study was performed at the Central Laboratory of Basic Medical College, Chongqing Medical University from January to October 2007. MATERIALS： Seventy-five female SD rats, weighing 210-230 g. rhEPO injection was provided by Sansheng pharmaceuticals company, Shenyang City, Liaoning Province, China, and the License number was HMLN S20010001. METHODS： A total of 75 female rats were randomly divided into rhEPO treatment, control, and sham operation groups, with 25 rats in each group. Rat models of facial nerve injury were established in the rhEPO treatment group and the control group by crushing the main trunk of the left facial nerve. Surgical microscopic observation of the facial nerve damage displayed perineurial disruption. The left stylomastoid foramen of the sham operation group were only exposed, but without nerve injury. The rhEPO treatment group was treated with rhEPO （5 000 U/kg, i.p.） once following injury and once a day for two weeks. The control and sham operation groups were treated with the same dose of normal saline （i.p.）, once following injury and once a day for two weeks. MAIN OUTCOME MEASURES： Rats were sacrificed 3, 7, 14, 21, and 28 days after injury, FMN survival after facial nerve injury was analyzed by Toluidine blue staining, and then survival ratios （L/R） were calculated. The number of apoptotic profiles in the injured FMNs were evaluated by TUNEL staining. Expression of Caspase-3 in the facial nucleus was detected by immunohistochemistry methods. RESULTS： A total of 75 rats were included in the final analysi     

Neuroprotective effects of recombinant human erythropoietin on facial motoneurons after facial nerve injury in rats

BACKGROUND: Erythropoietin and recombinant human erythropoietin (rhEPO) inhibit apoptosis of motor neurons caused by spinal cord injury and brain damage in rats. However, it still remains to be shown whether rhEPO can protect facial motoneurons (FMNs) as well.OBJECTIVE: To test the neuroprotective effects of rhEPO on injured FMNs, as well as the influence on Caspase-3 expression.DESIGN, TIME AND SETTING: Randomized, controlled, animal experiment. This study was performed at the Central Laboratory of Basic Medical College, Chongqing Medical University from January to October 2007.MATERIALS: Seventy-five female SD rats, weighing 210-230g. rhEPO injection was provided by Sansheng pharmaceuticals company, Shenyang City, Liaoning Province, China, and the License number was HMLN S20010001.METHODS: A total of 75 female rats were randomly divided into rhEPO treatment, control, and sham operation groups, with 25 rats in each group. Rat models of facial nerve injury were established in the rhEPO treatment group and the control group by crushing the main trunk of the left facial nerve. Surgical microscopic observation of the facial nerve damage displayed perineurial disruption. The left stylomastoid foramen of the sham operation group were only exposed, but without nerve injury. The rhEPO treatment group was treated with rhEPO (5000U/kg, i. p.) once following injury and once a day for two weeks. The control and sham operation groups were treated with the same dose of normal saline (i. p.), once following injury and once a day for two weeks.MAIN OUTCOME MEASURES: Rats were sacrificed 3, 7, 14, 21, and 28 days after injury, FMN survival after facial nerve injury was analyzed by Toluidine blue staining, and then survival ratios (L/R) were calculated. The number of apoptotic profiles in the injured FMNs were evaluated by TUNEL staining. Expression of Caspase-3 in the facial nucleus was detected by immunohistochemistry methods.RESULTS: A total of 75 rats were included in the final analysis. FMN survival ratios, both in rhEPO treatment group and control group, decreased gradually between seven and 28 days; however, FMN survival ratios were significantly greater in the rhEPO treatment group compared to the control group (P<0.05). No TUNEL-positive cells were observed three days after injury in the rhEPO treatment and control groups; however, by seven days after injury, apoptotic cells were observed and peaked by 14 days in the control group. Between seven and 21 days, apoptotic cell numbers were significantly lower in the rhEPO treatment group compared to the control group (P<0.05). The expression of Caspase-3 increased three days after injury and peaked at 14 days in the control group. Nevertheless, Caspase-3 expression was significantly lower in the rhEPO treatment group compared to the control group at each time point (P<0.05).CONCLUSION: Treatment with rhEPO can effectively protect facial motoneurons by reducing expression of Caspase-3 and inhibiting apoptosis.     

Skeletal muscle regeneration in young rats is dependent on growth hormone.

Skeletal muscle fibres have a well known ability to regenerate after different kinds of injury. This study was undertaken to establish if regenerating skeletal muscle is dependent on growth hormone (GH) in the same manner as normal, growing skeletal muscle in young rats. Muscle regeneration was achieved by injection of notexin into the soleus muscle. Initial necrosis, which included all muscle fibres, was followed by a rapid and uniform regeneration throughout the muscle. Cell proliferation was estimated by scintillation counting and autoradiography of incorporated [3H]thymidine, injected intravenously 1 h before killing, 7 or 27 days after the initiation of regeneration. GH deficiency was accomplished by hypophysectomy 4 days before the [3H]thymidine injection. Cell proliferation was diminished in both regenerating and normal muscle of the hypophysectomized rats compared to control and GH-substituted rats. After 7 days of regeneration the reduction of cell proliferation seen in hypophysectomized rats was less pronounced in the regenerating than in the normal muscle. These findings demonstrate that GH plays an important role for muscle regeneration in young rats, although other substances appear to be of greater importance during the early stages of regeneration.     

Skeletal muscle imaging in facioscapulohumeral muscular dystrophy,pattern and asymmetry of individual muscle involvement

To better understand postural and movement disabilities, the pattern of total body muscle fat infiltration was analyzed in a large group of patients with facioscapulohumeral muscular dystrophy. Additionally, we studied whether residual D4Z4 repeat array length adjusted for age and gender could predict the degree of muscle involvement. Total body computed tomography scans of 70 patients were used to assess the degree of fat infiltration of 42 muscles from neck to ankle level on a semi-quantitative scale. Groups of muscles that highly correlated regarding fat infiltration were identified using factor analysis. Linear regression analysis was performed using muscle fat infiltration as the dependent variable and D4Z4 repeat length and age as independent variables. A pattern of muscle fat infiltration in facioscapulohumeral muscular dystrophy could be constructed. Trunk muscles were most frequently affected. Of these, back extensors were more frequently affected than previously reported. Asymmetry in muscle involvement was seen in 45% of the muscles that were infiltrated with fat. The right-sided upper extremity showed significantly higher scores for fat infiltration compared to the left side, which could not be explained by handedness. It was possible to explain 29% of the fat infiltration based on D4Z4 repeat length, corrected for age and gender. Based on our results we conclude that frequent involvement of fat infiltration in back extensors, in addition to the abdominal muscles, emphasizes the extent of trunk involvement, which may have a profound impact on postural control even in otherwise mildly affected patients.     

Skeletal muscle NAD+(P) and NADP+-dependent malic enzyme in Friedreich's ataxia

Malic enzymes were studied in skeletal muscle from seven patients with Friedreich's ataxia (FA) and nine controls. Muscle contained three different malic enzymes. There were two strictly NADP+-dependent enzymes, one in the cytosol and one in mitochondria. These two enzymes are not allosteric. In FA muscle, activity of the mitochondrial NADP+-linked enzyme was significantly low and the cytosol NADP+-linked enzyme was significantly increased. A third malic enzyme, NAD+(P)-dependent, was found in the mitochondrial fraction. That enzyme had allosteric properties, and its activity was about the same in FA and control muscle.     

Mitochondrial impairment of human muscle in Friedreich ataxia in vivo

Friedreich ataxia occurs due to mutations in the gene encoding the mitochondrial protein frataxin. This (31)P magnetic resonance spectroscopy study on the calf muscle of Friedreich ataxia patients provides in vivo evidence of a severe impairment of mitochondrial function. Mitochondrial adenosine triphosphate resynthesis was studied by means of the post-exercise recovery of phosphocreatine. After ischemic exercise in calf muscles of all patients, phosphocreatine recovery was dramatically delayed. Time constants of recovery correlated with mutations of the frataxin gene, the age of the patients, and disease duration. (31)P magnetic resonance spectroscopy represents the first expedient tool for monitoring therapeutic trials in Friedreich ataxia non-invasively.     

重组人促红细胞生成素(rhEPO)对帕金森病大鼠模型小胶质细胞活化的影响

目的探讨重组人促红细胞生成素(recombinant human erythropoietin,rhEPO)对6-羟基多巴胺(6-OHDA)诱导的SD大鼠帕金森病(PD)模型小胶质细胞活化的影响。方法 40只SD大鼠随机分为A组(rhEPO+6-OHDA)、B组(生理盐水+6-OHDA)、C组(6-OHDA)、D组(生理盐水),每组10只。(1)A组:右侧纹状体内立体定向注射重组促红细胞生成素(rhEPO),24h后同侧黒质内立体定向注射6-OHDA;(2)B组:右侧纹状体内立体定向注射与rhEPO等量的生理盐水,24h后同侧黒质内立体定向注射6-OHDA;(3)C组:右侧黒质内立体定向注射6-OHDA;(4)D组:右侧黒质内立体定向注射与6-OHDA等量的生理盐水。4w后采用免疫组化检测黒质内酪氨酸羟化酶(TH)阳性神经元和CD11b阳性细胞数量及CD11b阳性细胞形态变化。结果与D组比较,A组大鼠黒质TH阳性神经元明显减少,CD11b阳性细胞明显增多,大部分小胶质细胞胞体小,突起细长;与B组和C组比较,A组大鼠黒质TH阳性神经元显著增多,CD11b阳性细胞显著减少,仅有少量小胶质细胞胞体大,突起短粗。结论重组人促红细胞生成素(rhEPO)可能通过抑制小胶质细胞活化,减轻6-OHDA对多巴胺(DA)能神经元的毒性损害,对DA能神经元产生神经保护作用。     

Neuroprotective effects of recombinant human erythropoietin in the developing brain of rat after lithium-pilocarpine induced status epilepticus

Status epilepticus triggers a mixture of apoptotic and necrotic cell death within the hippocampus. This neuronal loss may result in the development of epilepsy and cognitive deficits. Erythropoietin mediates a number of biological actions within the central nervous system and has been shown to be neuroprotective. In the present study, we investigated the effects of recombinant human erythropoietin on hippocampus of rat after lithium-pilocarpine induced status epilepticus. Twenty-one dam reared Wistar male rats, 21-day-old were divided into three groups: control group, lithium-pilocarpine induced status epilepticus and lithium-pilocarpine induced status epilepticus and erythropoietin treated group. Erythropoietin treated group received recombinant human erythropoietin 10 U/g intraperitoneally 40 min after pilocarpine injection for 5 days. Rats were sacrificed and brain tissues were collected at 5th day of experiment. Neuronal cell death and apoptosis were evaluated. Histopathological examination showed that erythropoietin significantly decreased neuronal cell death in CA1, CA2, CA3 and dentate gyrus regions of hippocampus. It also diminished apoptosis in the CA1 and dentate gyrus regions of hippocampus. In conclusion, erythropoietin may preserve the number of neurons and decrease apoptosis in model of status epilepticus induced by lithium-pilocarpine. This experimental study suggests that erythropoietin administration may be neuroprotective in status epilepticus.     

Skeletal muscle involvement in mucopolysaccharidosis type IIA: severe type of Hunter syndrome

A patient with mucopolysaccharidosis type IIA (MPS IIA) and progressive gait disturbance is described. The histopathology of biopsied muscle was studied; Dorling's method revealed muscle fibers and interstitial cells containing metachromatic granules which suggested the storage of sulfated acidic glycosaminoglycans. Electron microscopy demonstrated that the membrane-bound vacuoles were present in muscle fibers, subsarcolemmal area, vascular endothelial cells, satellite cells, and endomysial fibroblasts. Besides clinical features, this ultrastructural pathology in MPS IIA muscles of MPS IIA was more severe than MPS IIB muscles. The accumulation of glycosaminoglycans in muscle tissue may be an additional factor contributing to gradual motor impairment of patients with MPS IIA.     

实验性脑出血大鼠血肿周围VEGF、bFGF表达及血管新生的动态变化及重组人促红细胞生成素对其干预作用的研究

目的研究大鼠脑出血后血管内皮细胞生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)表达的动态变化及血管新生情况,以及重组人促红细胞生成素(rhEPO)对其的干预作用,探讨rhEPO对脑出血的可能保护机制。方法采用自体血脑内注入法建立脑出血动物模型。168只雄性SD大鼠随机分为正常组、假手术组、模型组、rhEPO治疗组。免疫组化法检测VEGF、bFGF、CD34的表达变化,用CD34的表达来反映血管新生情况。结果脑出血后3h即有VEGF表达增多,7～14d达高峰;bFGF的表达6h开始增多,72h达高峰;CD34+血管数12h开始增多,14d达高峰。rhEPO治疗组6h～21d时VEGF、bFGF表达比模型组高(P<0.05或P<0.01),且VEGF表达于72h～7d时提前达高峰;12h～21d时CD34+血管数高于模型组(P<0.01),且于7d时提前达高峰。结论rhEPO能上调脑出血后VEGF、bFGF的表达,促进脑出血后血肿周围新生血管生成。     

Improvement of cognitive functions in chronic schizophrenic patients by recombinant human erythropoietin

Schizophrenia is increasingly recognized as a neurodevelopmental disease with an additional degenerative component, comprising cognitive decline and loss of cortical gray matter. We hypothesized that a neuroprotective/neurotrophic add-on strategy, recombinant human erythropoietin (rhEPO) in addition to stable antipsychotic medication, may be able to improve cognitive function even in chronic schizophrenic patients. Therefore, we designed a double-blind, placebo-controlled, randomized, multicenter, proof-of-principle (phase II) study. This study had a total duration of 2 years and an individual duration of 12 weeks with an additional safety visit at 16 weeks. Chronic schizophrenic men (N=39) with defined cognitive deficit (>or=1 s.d. below normal in the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS)), stable medication and disease state, were treated for 3 months with a weekly short (15 min) intravenous infusion of 40,000 IU rhEPO (N=20) or placebo (N=19). Main outcome measure was schizophrenia-relevant cognitive function at week 12. The neuropsychological test set (RBANS subtests delayed memory, language-semantic fluency, attention and Wisconsin Card Sorting Test (WCST-64) - perseverative errors) was applied over 2 days at baseline, 2 weeks, 4 weeks and 12 weeks of study participation. Both placebo and rhEPO patients improved in all evaluated categories. Patients receiving rhEPO showed a significant improvement over placebo patients in schizophrenia-related cognitive performance (RBANS subtests, WCST-64), but no effects on psychopathology or social functioning. Also, a significant decline in serum levels of S100B, a glial damage marker, occurred upon rhEPO. The fact that rhEPO is the first compound to exert a selective and lasting beneficial effect on cognition should encourage new treatment strategies for schizophrenia.     

重组人红细胞生成素改善小鼠颅脑创伤后认知功能的机制探讨

研究背景颅脑创伤后的炎症反应可导致继发性脑损伤,引起神经元凋亡和神经功能缺损。重组人红细胞生成素(rhEPO)具有神经保护作用,我们采用液压打击动物模型,观察经rhEPO治疗后脑组织中性粒细胞变化和神经元凋亡情况,以探讨该药对颅脑创伤后神经功能保护的潜在作用机制。方法采用液压打击模型模拟颅脑创伤,分别于创伤后第1、3和7天观察不同处理组小鼠海马区髓过氧化物酶阳性中性粒细胞和Caspase-3阳性神经元表达变化;并于创伤后第7~11天进行Morris水迷宫实验,记录小鼠逃避潜伏期。结果与假手术组相比,伤后第1天小鼠海马区髓过氧化物酶阳性中性粒细胞数目即开始增加(均P=0.000),并于第3天达峰值水平(均P=0.000),至第7天时减少(均P=0.000);Caspase-3阳性神经元数目于伤后第1天开始逐渐增多(均P=0.000),至第7天达峰值水平(均P=0.000)。与生理盐水组相比,rhEPO组小鼠伤后第1~7天海马区髓过氧化物酶阳性中性粒细胞和Caspase-3阳性神经元数目呈逐渐减少趋势,不同观察时间点差异具有统计学意义(均P=0.000)。与生理盐水组相比,rhEPO组小鼠于Morris水迷宫实验训练第3天逃避潜伏期开始缩短(P=0.013),随着训练时间的延长,组间差异具有统计学意义(P=0.011,0.000)。结论液压打击后给予rhEPO可促进小鼠颅脑创伤后认知功能的恢复。其可能机制与抑制创伤后局部炎症反应、减少神经元凋亡、促进神经功能恢复有关。