首发儿童精神分裂症患者脑源性神经营养因子水平及神经元凋亡研究

目的探讨首发儿童精神分裂症患者脑源性神经营养因子(BDNF)、β-微管蛋白III(Tuj-1)、胱天蛋白酶3(Caspase-3)的变化,为揭示儿童精神分裂症的病因和发病机制提供参考。方法以2015年8月-2018年3月在大理州第二人民医院就诊的符合《精神障碍诊断与统计手册(第4版)》(DSM-IV)诊断标准的儿童精神分裂症患者(n=35)为研究组,同期在大理州第二人民医院的体检儿童中随机选取健康儿童为对照组(n=30)。通过实时荧光定量逆转录-聚合酶链反应(RT-PCR)测定两组儿童血液中BDNF、Caspase-3和Tuj-1的mRNA表达水平。结果与对照组相比,研究组BDNF和Tuj-1 mRNA表达低(P0.05或0.01),Caspase-3 mRNA表达高(P0.01)。结论儿童精神分裂症患者的BDNF mRNA和Tuj-1 mRNA表达少,Caspase-3 mRNA表达多,BDNF mRNA、Tuj-1 mRNA和Caspase-3 mRNA可能与儿童精神分裂症相关。     

丁苯酞对轻度血管性认知障碍患者执行功能的影响

目的 评估口服丁苯酞对血管性轻度认知障碍患者执行能力的影响。 方法 选择轻度血管性认知功能障碍患者48例，分为观察组24例，对照组24例。观察组服用丁苯 酞200 mg，每日3次，对照组服用维生素E 100 mg，每日3次，共90 d。比较两组治疗前及治疗后连线A 和连线B测试、数字广度顺背倒背、言语流畅性测验（verbal fluency test，VFT）、画钟测验量表评分， 评价丁苯酞对患者执行功能的影响。 结果 治疗前，两组认知和执行功能差异无统计学意义。治疗后，观察组连线A（[ 53.65±0.88）vs （56.22±0.87）]和连线B（[ 174.95±1.99）vs（177.41±1.44）]测试评分低于对照组，差异有统计学意义； 数字广度顺背倒背测试（[ 5.05±1.22）vs（4.59±0.33）]和VFT（[ 13.29±0.88）vs（12.43±1.02）]评分 高于对照组，差异有统计学意义；两组画钟测试差异无统计学意义。 结论 丁苯酞可以改善轻度血管性认知障碍患者执行功能。     

形觉剥夺性高度近视豚鼠巩膜形态改变及缺氧诱导因子-1α和氧自由基在高度近视中的作用

目的　观察形觉剥夺性高度近视（form deprivation high myopia，FDHM）豚鼠巩膜形态变化，探讨缺氧诱导因子-1α（hypoxia-inducible factor-1α，HIF-1α）及氧自由基在高度近视中的作用。方法　将豚鼠适应性饲养1周后，随机分为空白对照组（25只）和模型组（25只）。模型组豚鼠右眼行眼睑缝合，所有模型组豚鼠均选择右眼作为FDHM组，对侧眼为自身对照组。空白对照组豚鼠不做任何处理。于造模前及造模后8周采用检影镜测量屈光度，A超进行生物测量。形觉剥夺8周以后处死豚鼠，观察巩膜形态和超微结构的变化，测定巩膜HIF-1α相对表达量，超氧化物歧化酶（superoxide dismutase，SOD）活力及丙二醛（malondialdehyde，MDA）的含量。结果　豚鼠形觉剥夺8周以后，FDHM组屈光度从（+3.59±0.33）D变为（-7.96±0.55）D，明显高于空白对照组（+0.89±0.32）D、自身对照组（-0.55±0.49）D（均为P<0.05）；玻璃体腔深度为（4.12±0.13）mm明显高于空白对照组（3.71±0.23）mm和自身对照组（3.93±0.04）mm（均为P<0.05）；眼轴长度为（8.93±0.22）mm明显长于空白对照组（7.95±0.37）mm和自身对照组（8.01±0.15）mm（均为P<0.05）。巩膜组织明显变薄，细胞外基质增多，成纤维细胞密度降低，胶原纤维平均直径减小。FDHM组巩膜中HIF-1α相对表达量、MDA含量明显高于空白对照组和自身对照组，SOD活力明显低于空白对照组和自身对照组（均为P<0.05）。结论　形觉剥夺8周后，豚鼠FDHM眼近视度数明显增加，玻璃体腔深度增加，眼轴延长，巩膜形态发生病理性变化；HIF-1α、SOD、MDA可能参与了FDHM的形成。     

Responses of Types 1 and 2 Neovascularization in Age-Related Macular Degeneration to Anti-Vascular Endothelial Growth Factor Treatment: Optical Coherence Tomography Angiography Analysis

Purpose: To compare the responses of types 1 (sub-pigment epithelial) and 2 (subretinal) neovascularization in neovascular age-related macular degeneration (AMD) to anti-vascular endothelial growth factor (VEGF) treatment.

Methods: Fifty-five treatment-naïve neovascular AMD eyes (53 patients) were retrospectively included for analysis. All patients were treated with three loading injections of anti-VEGF agent, followed by further injections as required. The lesion size and vascular density of type 1 and 2 lesions before and after treatment for 12 months were analyzed using optical coherence tomography angiography (OCTA).

Results: The mean lesion size of the type 1 neovascularization group (42 eyes) showed no significant change from 2.12 ± 1.01 mm² at baseline to 2.08 ± 0.91 mm² at 12 months (P = .682). However, the mean lesion size of type 2 neovascularization significantly decreased from 1.23 ± 0.93 mm² at baseline to 0.79 ± 0.61 mm² at 12 months (P = .022). The proportion of eyes with lesion sizes that decreased by more than 40% from baseline was also significantly higher for the type 2 compared to the type 1 neovascularization group (46.2% versus 11.9%, P = .007). Vascular density showed no significant changes for both groups after treatment and showed no association with the change in lesion size. There was no significant difference between the groups in terms of visual acuity improvement.

Conclusion: OCTA analysis revealed different responses to anti-VEGF treatment depending on the location of neovascularization in neovascular AMD. Type 2 neovascularization was significantly regressed compared to type 1 neovascularization after anti-VEGF treatment. However, the changes in vascular density and visual outcome showed no significant differences between groups after 12 months of treatment.     

Adaptation of Vero cells to suspension growth for rabies virus production in different serum free media

Vero cells are nowadays widely used in the production of human vaccines. They are considered as one of the most productive and flexible continuous cell lines available for vaccine manufacturing. However, these cells are anchorage dependent, which greatly complicates upstream processing and process scale-up. Moreover, there is a recognized need to reduce the costs of vaccine manufacturing to develop vaccines that are affordable worldwide. The use of cell lines adapted to suspension growth contributes to reach this objective.The current work describes the adaptation of Vero cells to suspension culture in different serum free media according to multiple protocols based on subsequent passages. The best one that relies on cell adaption to IPT-AFM an in-house developed animal component free medium was then chosen for further studies. Besides, as aggregates have been observed, the improvement of IPT-AFM composition and mechanical dissociation were also investigated.In addition to IPT-AFM, three chemically defined media (CD293, Hycell CHO and CD-U5) and two serum free media (293SFMII and SFM4CHO) were tested to set up a serum free culture of the suspension-adapted Vero cells (VeroS) in shake flasks. Cell density levels higher than 2 × 10⁶ cells/mL were obtained in the assessed conditions. The results were comparable to those obtained in spinner culture of adherent Vero cells grown on Cytodex 1 microcarriers.Cell infection with LP-2061 rabies virus strain at an MOI (Multiplicity of Infection) of 0.1 and a cell density of 8 ± 0.5 × 10⁵ cells/mL resulted in a virus titer higher than 10⁷ FFU/mL in all media tested. Nevertheless, the highest titer equal to 5.2 ± 0.5 × 10⁷ FFU/mL, was achieved in IPT-AFM containing a reduced amount of Ca⁺⁺ and Mg⁺⁺. Our results demonstrate the suitability of the obtained VeroS cells to produce rabies virus at a high titer, and pave the way to develop VeroS cells bioreactor process for rabies vaccine production.     

A DNA vaccine expressing an optimized secreted FAPα induces enhanced anti-tumor activity by altering the tumor microenvironment in a murine model of breast cancer

Cancer-associated fibroblasts (CAFs), major components of the tumor microenvironment (TME), promote tumor growth and metastasis and inhibit the anti-tumor immune response. We previously constructed a DNA vaccine expressing human FAPα, which is highly expressed by CAFs, to target these cells in the TME, and observed limited anti-tumor effects in the 4T1 breast cancer model. When the treatment time was delayed until tumor nodes formed, the anti-tumor effect of the vaccine completely disappeared. In this study, to improve the safety and efficacy, we constructed a new FAPα-targeted vaccine containing only the extracellular domain of human FAPα with a tissue plasminogen activator signal sequence for enhanced antigen secretion and immunogenicity. The number of CAFs was more effectively reduced by CD8⁺ T cells induced by the new vaccine. This resulted in decreases in CCL2 and CXCL12 expression, leading to a significant decrease in the ratio of myeloid-derived suppressor cells in the TME. Moreover, when mice were treated after the establishment of tumors, the vaccine could still delay tumor growth. To facilitate the future application of the vaccine in clinical trials, we further optimized the gene codons and reduced the homology between the vaccine and the original sequence, which may be convenient for evaluating the vaccine distribution in the human body. These results indicated that the new FAPα-targeted vaccine expressing an optimized secreted human FAPα induced enhanced anti-tumor activity by reducing the number of FAPα⁺ CAFs and enhancing the recruitment of effector T cells in the 4T1 tumor model mice.     

Inter-ictal assay of peripheral circulating inflammatory mediators in migraine patients under adjunctive cervical non-invasive vagus nerve stimulation (nVNS): A proof-of-concept study

Objective

To assay peripheral inter-ictal cytokine serum levels and possible relations with non-invasive vagus nerve stimulation (nVNS) responsiveness in migraineurs.