【作者】 李宏；

【导师】 曾其毅；

【作者基本信息】 南方医科大学 ， 儿科学， 2013， 博士

【摘要】 研究背景脑性瘫痪(cerebral palsy, CP,简称脑瘫)是自受孕开始至婴儿期非进行性脑损伤和发育缺陷所导致的一组综合征,主要表现为运动障碍及姿势异常,可伴有或不伴有精神发育迟缓及癫痫,是儿童时期最常见的神经系统疾病之一,严重影响儿童健康及人口素质。随着围产医学的发展、产妇保健水平的提高,早产儿、低出生体重儿的存活率显著增加,与其相对的是,早产儿脑性瘫痪的发病率却呈上升趋势。2006年Kirby等对美国四个州8岁以内儿童脑性瘫痪进行调查统计,患病率为2.9-3.8%o,平均为3.3%o。1997-1998年,我国六省(区)1岁、2岁、3岁、4岁、5岁、6岁年龄组患病率分别为2.26‰、2.24‰、1.94‰、1.58‰、1.80‰、1.77‰,平均患病率为1.92‰。我国现有脑性瘫痪患儿400～500万,致残率为42%-45%,每年新增脑性瘫痪患儿3～4万,已成为新的临床、公共卫生和社会问题。目前各国对脑性瘫痪一般采取三级防治措施。一级防治是找出致病原因,避免脑性瘫痪的出现；二级防治是对脑性瘫痪患儿早期发现、早期干预,尽量减少或减轻后遗症；三级防治是对已经形成脑瘫的患儿坚持康复训练,尽可能地改善功能,最大限度地提高生存质量。脑性瘫痪防治的重点在于完善的一级防治及有效的二级防治手段。脑性瘫痪研究的重点集中在病因分析、致病机制以及修复机制三个方面。但其核心仍是发病机制的探索。脑性瘫痪的发病机制尚不完全清楚,是自受孕开始至婴儿期非进行性脑损伤和发育缺陷所导致的一组综合征。所有可能导致胎儿脑损伤的危险因素统称为脑瘫的高危因素。研究小儿脑性瘫痪病因及相关围产期高危因素,对降低该病的患病率、早期监测及干预、减少致残率、提高我国人口素质具有重要意义,这也是脑瘫一级防治的核心和根本。目前的资料显示,早产／低出生体重、新生儿窒息/HIE、新生儿高胆红素血症和宫内感染是最主要的围产高危因素。随着医疗技术的进步,尤其是产科及NICU水平的逐年提高,围产高危因素的构成也在不断发生变化。但目前尚缺乏长时间跨度的围产期高危因素变化的研究。本课题第一部分即对近10年来新入院脑瘫患儿的围产高危因素进行调查,试图找到近年来围产高危因素的构成及其变化,为我国脑瘫的一级防治提供临床证据。由于脑性瘫痪致病机制的不明确,由此造成脑瘫治疗方法的混乱和不确定,为脑瘫的二级防治带来了困难。目前,国内外尚无根治脑瘫的治疗方法。康复运动训练是得到公认的有效治疗和改善脑瘫症状的治疗方法之一,但多数的研究限于临床观察,对于运动训练起效的机理阐述较少。缺氧缺血性脑损伤(Hypoxic-ischemic brain damage,HIBD)是脑性瘫痪的主要致病机制之一,长期是脑瘫研究的热点。为了进一步深入探讨运动训练治疗脑瘫的机理,本课题第二部分选用新生大鼠HIBD模型,观察早期运动训练对新生大鼠缺氧缺血性脑损伤远期神经功能、病理改变、空间学习记忆能力、感觉运动功能、超微结构的影响。试图探讨早期运动训练对HIBD新生大鼠作用的机理,为早期运动训练治疗脑瘫提供实验依据。无论是一级还是二级防治,其根本仍在于明确脑瘫确切的致病机制。近年来关于脑瘫致病机制的许多研究证实,脑瘫具有明显的遗传易感性。在对瑞典1959-1970年脑性瘫痪患儿调查中发现足月儿偏瘫占60%、痉挛型双瘫占45%,早产儿痉挛型双瘫占32%,几乎全部单纯型共济失调都与遗传有关。研究发现肿瘤坏死因子α、甘露糖结合凝集素基因的多态性会增加脑性瘫痪的危险性。位于19号染色体19q132上的ApoE基因,此基因与神经细胞表面LDL和VLDL受体结合进入神经元细胞,对维持、修复神经细胞膜结构,维持神经递质和参与脑损伤后修复起到重要作用。携带ApoE E4基因或等位基因E2,发生脑性瘫痪的风险性较未携带者明显增加。ApoE4与认知相关,可造成海马萎缩及其连接性丧失,进而造成记忆障碍,影响逻辑能力导致智力低下；还可以导致胆碱能递质减少,进而出现精神运动视觉障碍。位于染色体2q37上的尿苷二磷酸葡萄糖醛酸转移酶(UGTIA1)基因突变是导致新生儿高胆红素血症和导致不随意运动型脑性瘫痪的重要因素之一。UGTIA1基因通过启动子与TATA结合调节转录起始DNA序列,而它突变可造成DNA转录起始频率和精确性异常。纯合子UGTIA1基因突变可影响葡萄糖醛酸转移酶的结构使酶活性下降,导致新生儿血清总胆红素显著升高。临床观察也发现,暴露在相同高危因素下的不同胎儿或新生儿,其临床结局差别甚大。这使我们想到,脑瘫的致病机制中,遗传因素及微观的生物学通路皆有可能起到非常重要的作用。脑瘫是一种严重影响人类健康和生命的复杂疾病。某些生物学通路在其疾病的发生、发展和转移的过程中发挥了关键作用。随着生物学实验技术的飞速发展,以基因芯片数据为代表的海量实验数据的产出,通过一系列算法和模型整合和分析实验数据,鉴定和模拟疾病相关的生物学通路,发现了很多重要的生物学结论,解释了遗传因素及微观的生物学通路在脑瘫致病中的作用。因此本课题第三部分选取了临床上最常见的脑瘫类型——早产儿脑室周围白质软化症(Periventricular leukomalacia, PVL)作为研究对象,通过对患儿和健康儿童血浆中microRNA基因芯片的表达差异进行分析,探讨microRNA在PVL发病中的作用,为进一步了解PVL的微观生物学通路发病机理,为将来PVL的宫内超早期诊断以及PVL的基因治疗探索出一条新的思路。第一部分摘要目的：比较十年来脑性瘫痪患儿相关高危因素构成,探索伴随医疗水平提高等社会大环境的变化下,脑性瘫痪高危因素的变化趋势,为脑性瘫痪的预防及治疗提供依据。方法：回顾性统计分析2003年至2012年间入院首次诊断为脑性瘫痪的病例中患儿的病例数、性别、年龄及高危因素等构成,并对各年份脑瘫患儿高危因素进行对比分析。结果：纳入共1040例病例,其中男性750例,女性290例,男女比例为2.59：1。患儿年龄<1岁147例,占14.1%；1-3岁440例,占42.3%；3～6岁294例,占28.3%；≥6岁159例,占15.3%；在10年脑瘫病例近40种高危因素中,早产仍是最重要的高危因素,其次是产时窒息、黄疸、胎膜早破及不良妊娠。10年间高危因素中产前、产时、产后因素构成有差异(χ2=72.234,P<0.05),宫内窘迫、产时窒息、产后窒息及不明确时间HIE的构成有差异(χ2=93.722,P<0.05)。结论：①本研究发现脑性瘫痪就诊患儿中存在明显的男女性别差异,男性多于女性；②患儿首次就诊年龄偏大,说明高危儿早期发育监测及早期干预工作依然存在较多困难,加强教育,提高对脑瘫的认识是关键；③产前因素所占比重有上升趋势,早产仍是最首位的高危因素,其次是产时窒息、黄疸、胎膜早破及不良妊娠。不同时间窒息中产前因素所占比例呈上升趋势,加强孕期保健,提高产科技术仍要作为目前脑性瘫痪防治的重点。第二部分摘要目的：探讨早期运动训练对新生大鼠缺氧缺血性脑损伤远期神经功能、病理改变、空间学习记忆能力、感觉运动功能、超微结构的影响。方法：90只7d龄SD大鼠按随机数字表法分为运动组、对照组和假手术组,每组各30只。运动组和对照组制备成新生儿缺氧缺血性脑损伤模型,且运动组于缺氧缺血性脑损伤后1周开始每天给予抓握、旋转、行走、平衡等训练。4周后进行神经功能评分,尼氏染色计数海马CA1区和皮层额部神经元数量,并检测突触素、c-fos表达水平；水迷宫检测各组大鼠空间学习记忆能力和感觉功能；透射电镜观察突触和神经元超微结构。结果：运动组大鼠神经功能评分、空间学习记忆能力均优于对照组,差异有统计学意义(均P<0.05)。对照组大鼠左侧海马CA1区及皮层额部尼氏染色阳性神经元数目较运动组和假手术组明显减少,差异有统计学意义(F=45.550,P<0.05；F=82.521,P<0.05)。运动组大鼠海马CA1和皮层额部突触素、c-fos表达水平明显高于对照组,差异有统计学意义(突触素：F=81.747, P<0.05; F=26.865, P<0.05。c-fos:F=680.750, P<0.05; F=296.617, P<0.05)。透射电镜下显示对照组大鼠海马和皮层突触损伤严重,神经毡区突触减少,突触前膨大肿胀、轮廓不清晰,突触小泡溶解、空泡形成,突触后致密区变薄、薄厚不均,而运动组未见明显异常。结论：早期运动训练可减少缺氧缺血性脑损伤后海马和皮层额部神经元损伤,增强突触可塑性,从而改善缺氧缺血性脑损伤后远期神经功能,其中突触素和c-fos在海马和皮层额部表达增强可能是其改善的机制之一。第三部分摘要目的：比较脑室周围白质软化(periventricular leukomalacia, PVL)患儿和健康儿童血清中microRNA基因芯片的差异表达,筛选出显著差异表达谱,为PVL早期发现、早期诊断提供生物学标记物。并为下一步探讨microRNA在PVL疾病中的发病机制,提供一条新的思路。方法：收集7例PVL患儿和5例健康儿童的全血,2000r/min离心10min分离血清,从血清中提取RNA,使用Exiqon公司的microRNA芯片筛选microRNA表达谱。以microRNA芯片表达结果为基础,采用microcosm, miranda和targetscan三个数据库及GO分析,筛选出与神经、血管、遗传发育靶基因相关的10个microRNA:hsa-miR-133b、hsa-miR-323a-3p、hsa-miR-330-3p、 hsa-miR-654-5p、hsa-miR-498、hsa-miR-574-5p、hsa-miR-638、hsa-miR-149-3p、 hsa-miR-1228-5p、hsa-miR-671-3p,对其进行实时荧光定量PCR验证。结果：PVL患儿中microRNA表达谱与健康患儿相比较有显著差异,在3100个microRNA中,有100个microRNA存在差异表达,34个microRNA表达上调,66个表达下调。用实时荧光定量PCR技术验证10个筛选出的microRNA,其中hsa-miR-323a-3p、hsa-miR-671-3p、hsa-miR-330-3p、hsa-miR-654-5p、 hsa-miR-498、hsa-miR-149-3p、hsa-miR-1228-5p共7个与microRNA芯片结果具有同样趋势。结论：部分microRNA在PVL疾病中的表达有显著差异,提示microRNA有可能与PVL的发病相关,为今后PVL的早期诊断、生物学机制探讨和基因治疗提供新的思路。全文结论1.脑性瘫痪就诊患儿中存在明显的男女性别差异；患儿就诊年龄偏大；10年间产前因素所占比重有上升趋势,不同时间窒息中产前因素所占比例呈上升趋势。提示,加强孕期保健及胎儿发育监测、提高产科技术依然是目前脑性瘫痪早期防治的重点。2.早期运动训练可减少缺氧缺血性脑损伤后海马和皮层额部神经元损伤,增强突触可塑性,从而改善缺氧缺血性脑损伤后远期神经功能,其中突触素和c-fos在海马和皮层额部表达增强可能是其改善的机制之一。3.部分microRNA在PVL疾病中的表达有显著差异,提示microRNA有可能与PVL的发病相关,为今后PVL的早期诊断、生物学机制探讨和基因治疗提供新的思路。展望1.扩大PVL病例数(100例)进一步验证microRNA芯片结果准确性,从而作为PVL早期诊断的生物学标记物。2. microRNA在PVL发病中关于神经、血管、遗传发育等生物学通路中的作用,是我们今后探讨的重点,希望能从微观角度挖掘其致病的分子机理。3.早期功能训练结合基因工程干预(上调或下调相关的microRNA,改变其生物学特征,为临床治疗服务)是今后治疗脑瘫患儿的一个富有希望的方向。更多还原

【Abstract】 Research backgroundCerebral palsy is a group of syndromes,caused by non-progressive brain injury and developmental defects which occured from the moment of conception to infancy.The main clinical manifestations are movement disorders and posture abnormalities, may be associated or not associated with mental retardation and epilepsy.It is one of the most common neurological diseases of childhood andhas a serious impact on children’s health and the quality of the population.With the development of perinatal medicine and the elevation of maternal health level,the survival rate of premature children and low birth weight children got a significant upgrade,relatively,the incidence of preterm children with cerebral palsy showed an upward trend.Kirby conducted population-based surveillance of8-year-old children in2006, in four states of America, found the CP prevalence was3.3per1000, ranging from2.9to3.8per1000.In1997-1998, a surveyof China’s six provinces (regions) child1year old,2years old,3years old,4years old,5years old,6-year-old age group prevalence rates were2.26per1000、2.24per1000、1.94per1000、1.58per1000、1.80per1000、1.77per1000, the average prevalence rate1.92per1000。The number of children with cerebral palsy in China is from4to5billion, the morbidity is for42percent to45percent, thus an annual increase of cerebral palsy children is about30million to40million. This has become a new clinical problems, public health problems and social problems. At present we general treat cerebral palsy with three-level prevention and control measures. Primary prevention is to find out the causes, to avoid the occurrence of cerebral palsy; Secondary prevention is early discovery, early intervention, as far as possible to reduce or relieve sequelae; Tertiary prevention is help the children insist to cerebral palsy rehabilitation training, as much as possible to improve function, maximize quality of life. The key of prevention and control of cerebral palsy is to perfect the primary prevention and secondary prevention measures. The researches about cerebral palsy focused on the cause analysis, pathogenic mechanism and repair mechanism. However, the core is still the pathogenesis exploration.The pathogenesis of cerebral palsy is not entirely clear,since it is a group of syndromes of non-progressive brain injury and developmental defects caused from the moment of conception to infancy. All risk factors that may cause fetal brain damage collectively referred to as risk factors for cerebral palsy. Study of cerebral palsy causes and perinatal risk factors, to reduce the prevalence of the disease, early detection and intervention to reduce morbidity and improve the quality of our population is of great significance, which is the core of cerebral palsy primary prevention and fundamental. The current data show that preterm/low birth weight, neonatal asphyxia/HIE, neonatal hyperbilirubinemia and intrauterine infection is the most important perinatal risk factors. With the advances in medical technology, especially the obstetric and NICU level increase year-on-year, perinatal risk factors constitute also constantly change. However, there is a lack of researches on the change of long time span of perinatal high-risk factors. The first chapter of this subject that survey the newly admitted children with cerebral palsy perinatal risk factors in the past10years, trying to find the formation and change of perinatal risk factors in recent years, provide clinical evidence for cerebral palsy of primary prevention in China.Due to the pathogenesis of cerebral palsy is not clear, causing confusion and uncertainty to the cerebral palsy treatment, makes it difficult for the secondary prevention and therapy of cerebral palsy. At present, there is no radical treatment of cerebral palsy, including domestic and overseas. Rehabilitation exercise training is recognized one of the effective treatment and improve the symptoms of cerebral palsy, but most of the research is limited to clinical observation, and less elaborated mechanism for sports training onset. Hypoxic-ischemic brain damage (HIBD) is one of the main pathogenic mechanisms of cerebral palsy, and long-term cerebral palsy research hotspot. In order to further investigate the mechanism of exercise training in the treatment of cerebral palsy, the second chapter in this study selected neonatal rat HIBD model to observe long-term neurological function, pathological change, spatial learning and memory, sensory and motor function and ultrastructure after the early exercise training on hypoxic-ischemic brain damage. An attempt was made to explore the mechanism of early exercise training on HIBD in neonatal rats to provide experimental evidence for the early exercise training in the treatment of cerebral palsy.Either primary or secondary prevention, the fundamental is remain the definite pathogenic mechanism of cerebral palsy. In recent years, many studies on the pathogenesis of cerebral palsy show that cerebral palsy has obvious genetic predisposition. Full-term children with hemiplegia accounted for60%, spastic diplegia (45%), preterm children with spastic diplegia32%[2] found in children with cerebral palsy in Sweden from1959to1970survey, almost all pure Freemasonry disorders related to heredity. Studies found that the gene polymorphisms tumor necrosis factor alpha and mannose-binding lectin will increase the risk of cerebral palsy. ApoE gene, located on chromosome1919q132, incorporated into neuronal cell throught binding the surface receptor LDL and VLDL of nerve cells, play an important role in maintaining and repairing nerve cell membrane structure, as well as maintaining neurotransmitter involved in brain damage and repair. Carry the ApoE E4genes or alleles E2significantly increased the risk of cerebral palsy compared with noncarriers. ApoE4is associated with cognitive, can cause hippocampal atrophy and its loss of connectivity, which causes memory impairment, affect the logical ability leads to mental retardation; also can cause a decrease in cholinergic neurotransmitters, thus lead to psychomotor visually impaired. Located on chromosome2q37uridine diphosphate glucuronyl transferase (UGT1A1) gene mutations is one important factor of resulting in neonatal hyperbilirubinemia and leading to dyskinetic cerebral palsy. UGT1A1gene via promoter binding TATA to regulate transcription initiation DNA sequences, which mutations can cause abnormal frequency and accuracy of DNA transcription initiation. The homozygous UGT1A1gene mutations can affect the structure of the glucuronyl transferase enzyme and decrease the activity, resulting significantly higher in neonatal serum total bilirubin.Clinical observation also found that different fetal or neonatal exposure to the same risk factors got great different clinical outcome. This give us some suggestions-genetic factors would play an important role in pathogenic mechanism of cerebral palsy. In order to understand the role of genetic factors in pathogenic mechanism of cerebral palsy, we selected the most common clinically type of cerebral palsy-premature with periventricular leukomalacia (PVL) as the study object in the third chapter of this subject. By way of comparative analysis microRNA expressions in plasma of children patients and healthy children, to explore the role of microRNAs in the pathogenesis of PVL, get further understand of the pathogenesis of PVL, then explore a new way for the early diagnosis of intrauterine PVL and gene therapy of PVL.Abstract of part oneObjective To compare the related risk factors of cerebral palsy children in ten years, explore the trend of cerebral palsy risk factors with the change of social environment such as medical level and provide the basis for the prevention and treatment of cerebral palsy.Methods The hospitalized cases of cerebral palsy diagnosed for the first time in ZhuJiang Hospital of Southern Medical University from2003to2012were analyzed retrospectively, with the constituent of the number, gender, age and risk factors in each year.Results There are1040cases in this study. There were750males and290females giving a male female ratio of2.59:1. Children<1year old in147cases, accounting for14.1%;1year old-3years old in440cases, accounting for42.3%;3year old-6years old in294cases, accounting for28.3%,>6years old in159cases, accounting for15.3%. In nearly40kinds of risk factors of cerebral palsy of10years, preterm birth remains the most important risk factors, followed by the intrapartum asphyxia, jaundice, adverse pregnancy, and premature rupture of membranes. The constituent of antenatal, intrapartum and postpartum factors were different (χ2=72.234, P<0.05). The constituent of intrauterine distress, intrapartum asphyxia, postpartum asphyxia and not clear time HIE were difference (χ2=93.722, P<0.05).Conclusions (1) This study found that there is significant gender difference in cerebral palsy children, male more than female;(2) Age of first diagnosed with cerebral palsy is large, which shows that early development monitoring and early intervention for high-risk children is remain difficult, and it is important to strengthen education and to raise awareness of cerebral palsy;(3) Prenatal factors have a rising trend, preterm birth remains the most important risk factors, followed by the intrapartum asphyxia, jaundice, adverse pregnancy, and premature rupture of membranes. The proportion of prenatal factors in the different time asphyxia is on the rise, to strengthen pregnancy care and to improve obstetric techniques are still as is currently the focus of prevention and cure of cerebral palsy.Abstract of part twoObjective To investigate the effect of early physical training on long-lasting neurological,pathology,spatial learning ability,sensorimotor functionand ultrastructure in neonatal rats submitted to hypoxic-ischemic brain damage(HIBD).Methods ninety7-day-old Sprague-Dawley rats were randomly divided into three groups:a group that was subjected to left carotid ligation followed by2hours hypoxic stress(the control group);a group that received physical training(Grab,Rotation,Walk,balance)1weeks after the HIBD event(the trained group);a sham-operation group that was subjected to a sham-operation without ligation and hypoxic stress(the sham-operation group.Following four weeks physical training, neurological score, the expression levels of synaptophysin and c-fos were examined; Morris water maze tests and cortex sensorimotor tests were performed; Left hippocampal CA1and cortex neurons and ultrastructure were performed.Results Compared with the control group, the Neurological score、Spatial learning and memory ability and sensorimotor tests of the trained group were significantly increased, whereas there was no significant difference between the trained group and the sham-operation group. The neurons in the left hippocampal CA1zone and cortex were decreased of the control group.The significant difference compared with trained group was obvious(F=45.550, P<0.05; F=82.521, P<0.05). The expression of synaptophysin and c-fos in the trained group increased significantly compared with that in the control group(synaptophysin:F=81.747, P<0.05; F=26.865, P<0.05. c-fos:F=680.750, P<0.05; F=296.617,P<0.05).The ultrastructure of the left hippocampus and cortex was remarkably abnormal in the control group by the transmission electron microscopy, while no obvious abnormality was observed in the trained group and the sham-operation group.Conclusions Early physical training can restrain brain damage and ameliorate spatial learning and memory impairments in rats with HIBD. Early exercise rehabilitation can reduce hippocampal and cortical neuronal damage, enhanced synaptic plasticity,and ameliorate the long-term neurological function after HIBD.The strong expression of synaptophysin and c-fos in the hippocampus and cortex caused by early physical training may be one of the improvement mechanisms.Abstract of part threeObjectiveTo study the differential microRNAs experssion between patients with periventricular leukomalacia(PVL) and healthy controls, filter out thesignificant differences inexpression profilesforthePVLearlydetection, early diagnosisbiological markers. And further moretoinvestigatemicroRNAsinthe pathogenesis ofPVLdisease, provideanew way of thinking.Methods Whole blood from7PVL patients and5controls healthies were separated into plasma at2000rpm for10minutes. RNA were harvested using kit.MicroRNAs profiling were performed using Exiqon microRCURY LNA microRNAs array.In addition, Using microcosm,miranda,targetscanand GO analysis, to screen10microRNAs,including hsa-miR-133b, hsa-miR-323a-3p, hsa-miR-330-3p, hsa-miR-654-5p, hsa-miR-498, hsa-miR-574-5p, hsa-miR-638, hsa-miR-149-3p, hsa-miR-1228-5p, hsa-miR-671-3p, associated with the nerves, blood vessels, genetic development of target genes.Then validating them through Real-time quantitative PCR.ResultsMicroRNAs expression profile was found to be differentially in the PVL patients compared with the healthy donors. Of3100microRNAs detected on the microarray,100microRNAs were found to be differentially expressed in the PVL patients,34pieces of microRNA were up-regulated more than twice in the PVL patients,66pieces were down-redulated more than twice. Real-time PCR detected10microRNAs:hsa-miR-323a-3p, hsa-miR-671-3p, hsa-miR-330-3p, hsa-miR-654-5p, hsa-miR-498, hsa-miR-149-3p, hsa-miR-1228-5p,7of them have the same trend with microRNA microarray results.ConclusionsThere are significant differences of microRNAs between PVL patients and healthy donors, it maybe play an important role in pathogenesis of PVL. We hope to provide new ideasforthefutureof PVLearly diagnosis, biological mechanismsandgene therapy.Full text Conclusions1. Obviously, the gender of the children with cerebral palsy is different. The proportion of older children is large. Prenatal factors have a rising trend, and the proportion of prenatal factors in the different time asphyxia is on the rise. These findings suggest that strengthen prenatal care and fetal growth monitoring, improve obstetric technology are still the focus of early prevention and treatment of cerebral palsy.2. Early physical training can restrain brain damage and ameliorate spatial learning and memory impairments in rats with HIBD. Early exercise rehabilitation can reduce hippocampal and cortical neuronal damage, enhanced synaptic plasticity,and ameliorate the long-term neurological function after HIBD.The strong expression of synaptophysin and c-fos in the hippocampus and cortex caused by early physical training may be one of the improvement mechanisms.3. There are significant differences of microRNAs between PVL patients and healthy donors, it maybe play an important role in pathogenesis and early diagnosis of PVL.Prospect1. Expanding the number of PVL cases (n=100) to further verify the accuracy of the microRNA microarray results, thus as biomarkers of PVL early diagnostic.2. Our direction is to specific the role of microRNA in the incidence of PVL about nerves,blood vessels, and genetic pathways,and to further discuss from the microscopic view.3. Early functional training combined with genetic engineering intervention (Upgrade or downgrade the mirna to change its biological characteristics, so that can service for clinical treatment)provide a promising future treatment of children with cerebral palsy.更多还原