Co-editing PINK1 and DJ-1 Genes Via Adeno-Associated Virus-Delivered CRISPR/Cas9 System in Adult Monkey Brain Elicits Classical Parkinsonian Phenotype

Whether direct manipulation of Parkinson’s disease (PD) risk genes in the adult monkey brain can elicit a Parkinsonian phenotype remains an unsolved issue. Here, we used an adeno-associated virus serotype 9 (AAV9)-delivered CRISPR/Cas9 system to directly co-edit PINK1 and DJ-1 genes in the substantia nigras (SNs) of two monkey groups: an old group and a middle-aged group. After the operation, the old group exhibited all the classic PD symptoms, including bradykinesia, tremor, and postural instability, accompanied by key pathological hallmarks of PD, such as severe nigral dopaminergic neuron loss (>64%) and evident α-synuclein pathology in the gene-edited SN. In contrast, the phenotype of their middle-aged counterparts, which also showed clear PD symptoms and pathological hallmarks, were less severe. In addition to the higher final total PD scores and more severe pathological changes, the old group were also more susceptible to gene editing by showing a faster process of PD progression. These results suggested that both genetic and aging factors played important roles in the development of PD in the monkeys. Taken together, this system can effectively develop a large number of genetically-edited PD monkeys in a short time (6–10 months), and thus provides a practical transgenic monkey model for future PD studies.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12264-021-00732-6.     

卵巢上皮肿瘤组织Fas、Angs、E-cad的表达水平及意义分析

目的检测Fas、促血管生成素-1(Ang-1)、促血管生成素-2(Ang-2)、E-钙粘蛋白(E-cad)在卵巢上皮肿瘤组织中的表达水平,藉此评价其与肿瘤发生发展及预后的关系。方法应用免疫组织化学S-P法和原位杂交技术,检查了21例卵巢上皮恶性肿瘤,18例良性肿瘤,8例正常卵巢组织中Fas,Ang-1,Ang-2及E-cad的表达。结果 Fas的表达随肿瘤病变程度的增高而逐步减弱,恶性肿瘤组织与正常卵巢及良性肿瘤组织间有显著差异(P<0．05);E-cad免疫反应阳性强度在卵巢上皮癌组织中明显降低,与正常卵巢及良性肿瘤组织间有显著性差异(P<0．05)。原位杂交显示,Ang-1 mRNA在正常卵巢及肿瘤组织中都有表达,其表达水平没有显著差异(P>0．05);Ang-2 mRNA在恶性肿瘤组织中表达明显上调(P<0．01);E-cad mRNA在恶性肿瘤组织中表达明显下调(P<0.01)。结论 Fas的表达水平与卵巢上皮肿瘤的内在发展相关;由E-cad介导的细胞间黏附作用的减弱是卵巢癌发生、发展及转移的一个重要因素;Ang-2对于促进卵巢癌组织血管新生和形成可能具有重要促进作用。     

Distinct Defects in Spine Formation or Pruning in Two Gene Duplication Mouse Models of Autism

Autism spectrum disorder (ASD) encompasses a complex set of developmental neurological disorders, characterized by deficits in social communication and excessive repetitive behaviors. In recent years, ASD is increasingly being considered as a disease of the synapse. One main type of genetic aberration leading to ASD is gene duplication, and several mouse models have been generated mimicking these mutations. Here, we studied the effects of MECP2 duplication and human chromosome 15q11-13 duplication on synaptic development and neural circuit wiring in the mouse sensory cortices. We showed that mice carrying MECP2 duplication had specific defects in spine pruning, while the 15q11-13 duplication mouse model had impaired spine formation. Our results demonstrate that spine pathology varies significantly between autism models and that distinct aspects of neural circuit development may be targeted in different ASD mutations. Our results further underscore the importance of gene dosage in normal development and function of the brain.     

Constitutive Descriptions and Restoration Mechanisms of a Fe-17Cr Alloy during Deformation at Temperatures of 700–1000 °C

The deformation behavior for highly purified Fe-17Cr alloy was investigated at 700~1000 °C and 0.5~10 s⁻¹. The microstructure evolution and corresponding mechanism during deformation were studied in-depth, using electron backscattering diffraction, transmission electron microscopy and precession electron diffraction. During deformation, dynamic recrystallization (DRX) occurred, along with extensive dynamic recovery, and the active DRX mechanism depended on deformation conditions. At higher Zener-Hollomon parameter (Z ≥ 5.93 × 10²⁷ s⁻¹), the development of the shear band was promoted, and then continuous DRX was induced by the formation and intersection shear band. At lower Zener-Hollomon parameter (Z ≤ 3.10 × 10²⁵ s⁻¹), the nucleation of the new grain was attributed to the combination of continuous DRX by uniform increase in misorientation between subgrains and discontinuous DRX by grain boundary bulging, and with increasing temperature, the effect of the former became weaker, whereas the effect of the latter became stronger. The DRX grain size increased with the temperature. For alleviating ridging, it seems advantageous to activate the continuous DRX induced by shear band through hot deformation with higher Z. In addition, the modified Johnson-Cook and Arrhenius-type models by conventional way were developed, and the modified Johnson-Cook model was developed, using the proposed way, by considering strain dependency of the material parameters. The Arrhenius-type model was also modified by using the proposed way, through distinguishing stress levels for acquiring partial parameter and through employing peak stress to determine the activation energy and considering strain dependency of only other parameters for compensating strain. According to our comparative analyses, the modified Arrhenius-type model by the proposed approach, which is suggested to model hot-deformation behavior for metals having only ferrite, could offer a more accurate prediction of flow behavior as compared to other developed models.     

舌下微循环变化在猪失血严重程度评估中的应用价值

目的探讨舌下微循环变化在猪失血严重程度评估中的应用价值。 方法健康雄性白猪20只,实验猪的总血容量被评估为70 ml/kg,根据失血比例的不同分为4组：对照组、25%失血组、35%失血组和45%失血组,每组5只。比较各组动物失血前体质量、心率、平均动脉压、心输出量、血乳酸及微血管流动指数（MFI）和灌注血管密度（PVD）变化,并分别于失血后30 min、1 h、2 h、3 h、4 h,评估动物的平均动脉压、心输出量、血乳酸、MFI及PVD的变化。 结果各组动物失血前体质量、心率、平均动脉压、心输出量、血乳酸、MFI及PVD比较,差异均无统计学意义（F = 0.156、0.260、0.467、0.417、0.434、1.778、0.149,P均> 0.05）。随着各组动物失血量的增加,平均动脉压及心输出量于失血后30 min[（117 ± 8）、（56 ± 10）、（45 ± 4）、（31 ± 5）mmHg,（4.80 ± 0.54）、（3.32 ± 0.32）、（2.35 ± 0.37）、（1.67 ± 0.15）L/min]及失血后1 h [（123 ± 4）、（89 ± 6）、（67 ± 8）、（52 ± 8）mmHg,（4.79 ± 0.63）、（3.99 ± 0.37）、（2.75 ± 0.35）、（2.05 ± 0.39）L/min]逐渐下降（P均< 0.05）,血乳酸于失血后1 h逐渐增加[（1.12 ± 0.33）、（2.10 ± 0.25）、（3.74 ± 1.38）、（5.88 ± 1.48）mmol/L,P均< 0.05],MFI[（2.92 ± 0.09）、（2.47 ± 0.28）、（1.30 ± 0.14）、（0.87 ± 0.27）,P均< 0.05]及PVD水平[（4.95 ± 0.12）、（4.20 ± 0.54）、（2.98 ± 0.10）、（2.64 ± 0.31）,P均< 0.05]于失血后30 min逐渐下降。Pearson相关性分析显示MFI及PVD与平均动脉压（r = 0.903、0.923,P均< 0.05）、心输出量（r = 0.919、0.919,P均< 0.05）呈显著正相关,与乳酸水平呈显著负相关（r = -0.706、-0.745,P均< 0.05）。 结论舌下微循环监测可实时反映机体失血程度,有助于判断失血严重程度。     

循环肿瘤细胞检测技术操作过程中存在的问题及对策

循环肿瘤细胞(CTC)是从恶性肿瘤组织中脱落,进入外周血循环的肿瘤细胞.CTC检测对于癌症的早期诊断,以及检测患者的术后复发与转移、评估疗效与预后、选择合适的个体化治疗等方面具有重要的临床意义,且能帮助理解CTC在外周血中的存活、吸附、侵袭等生物学行为,破译肿瘤转移的机制[1].一张合格的CTC制片是保证信号判读的前提...     

杜仲水提物对糖尿病大鼠血清和阴茎组织睾酮的影响

目的:研究杜仲水提物对糖尿病(DM)模型大鼠血清和阴茎组织睾酮(T)水平的影响.方法:采用四氧嘧啶两次给药法建立DM模型;30只DM大鼠随机分为赋形剂组(n=10)、西地那非组(n=10)和杜仲组(n=10).灌胃4周后,用放射免疫法检测血清和阴茎组织匀浆液中的睾酮水平;结果:杜仲组血清和阴茎组织睾酮水平显著高于赋形剂组(P＜0.01)和西地那非组(P＜0.01).结论:杜仲可提高DM大鼠血清和阴茎组织睾酮水平.     

鼓窦径路手术治疗先天性外耳道闭锁中耳畸形——附89例临床分析

目的 :探讨鼓窦径路治疗先天性外耳道闭锁中耳畸形的效果及重建耳道再闭锁的预防方法。方法 :回顾性分析 1993～ 1998年 5年间收治的 89例 (94耳 )先天性外耳道闭锁中耳畸形病人 ,均采用鼓窦径路重建外耳道 ,鼓室成形术 ,其中 型 80耳、加高 型 11耳、内耳开窗 3耳。康宁克通 A注射、扩张子扩张等预防耳道再闭锁。结果 :94耳均顺利找到鼓窦 ,开放鼓室 ,行听力重建 ,术后语言频率平均听力提高 2 0 d B以上者 80耳 (85 .1% ) ,提高 2 5 d B以上者 49耳(5 2 .1% )。再造耳道有闭锁征象者经康宁克通 A注射、扩张子扩张取得良好效果。结论 :鼓窦径路治疗先天性外耳道闭锁具有易掌握、安全、省时、效果可靠等特点 ,再造耳道有闭锁征象时应及时处理。