FasL gene knock-down therapy enhances the antiglioma immune response

Malignant glioma is a lethal form of brain cancer that is very difficult to treat. The aggressive behavior of these neoplasms and their limited responsiveness to therapy has been attributed in part to the ability of these tumors to evade the immune system. Gliomas, like many other solid tumors, express components of numerous immune escape mechanisms, including immunosuppressive proteins such as TGF-β, IL-10, and FasL. Here, we show that FasL expression can support the growth of experimental intracranial glioma. We show that FasL is readily detected in human glioblastoma multiforme clinical specimens. FasL was found to be expressed by three well-characterized rat glioma cell lines (9L, F98, and C6) and glioma cell-derived FasL mediated the death of phytohemagglutinin-stimulated Jurkat T-lymphocytes when cocultured with glioma cells in vitro. We asked if inhibiting 9L-derived FasL altered the growth of experimental glioma. FasL expression knockdown using shRNA reduced the growth of subcutaneous and intracranial 9L gliomas by ∼50% in immune competent Fisher 344 rats. In contrast, FasL expression knockdown had no affect on the growth of intracranial 9L glioma in T-cell deficient athymic rats. Intracranial tumors derived from FasL knockdown 9L glioma cells contained up to 3-fold more tumor infiltrating T-cells than tumors derived from control 9L cells. These results demonstrate that down-regulating FasL expression and/or function in glial malignancies can enhance T-cell tumor infiltration and inhibit tumor growth. The findings suggest that targeting endogenous FasL in glial malignancies could enhance the efficacy of emerging immune-based treatment strategies.     

Protective effects of Bushen Tiansui decoction on hippocampal synapses in a rat model of Alzheimer's disease

Bushen Tiansui decoction is composed of six traditional Chinese medicines:Herba Epimedii,Radix Polygoni multiflori,Plastrum testudinis,Fossilia Ossis Mastodi,Radix Polygalae,and Rhizoma Acorus tatarinowii.Because Bushen Tiansui decoction is effective against amyloid beta(Aβ) toxicity,we hypothesized that it would reduce hippocampal synaptic damage and improve cognitive function in Alzheimer's disease.To test this hypothesis,we used a previously established animal model of Alzheimer's disease,that is,microinjection of aggregated Aβ25–35 into the bilateral brain ventricles of Sprague-Dawley rats.We found that long-term(28 days) oral administration of Bushen Tiansui decoction(0.563,1.688,and 3.375 g/m L;4 m L/day) prevented synaptic loss in the hippocampus and increased the expression levels of synaptic proteins,including postsynaptic density protein 95,the N-methyl-D-aspartate receptor 2 B subunit,and Shank1.These results suggested that Bushen Tiansui decoction can protect synapses by maintaining the expression of these synaptic proteins.Bushen Tiansui decoction also ameliorated measures reflecting spatial learning and memory deficits that were observed in the Morris water maze(i.e.,increased the number of platform crossings and the amount of time spent in the target quadrant and decreased escape latency) following intraventricular injections of aggregated Aβ25–35 compared with those measures in untreated Aβ_(25–35)-injected rats.Overall,these results provided evidence that further studies on the prevention and treatment of dementia with this traditional Chinese medicine are warranted.     

过表达miR-126对人肺癌细胞体外生长的抑制作用和初步机制研究

目的:探讨过表达miR-126对人肺癌95D细胞体外生长的影响.方法:利用PCR技术成功构建pcDNA3.1(-)-pri-miR-126载体(命名为p-miR-126),将p-miR-126重组质粒体外瞬时转染人肺癌95D细胞,TaqMan探针法检测miR-126的相对表达水平,CCK-8(cell counting kit-8)法和划痕法分别检测95D细胞的增殖、迁移能力.Western Blot检测95D细胞中蛋白AKT和磷酸化AKT(phosphorylation AKT,p-AKT)的表达情况.结果:成功构建miR-126真核表达载体,且该载体可上调miR-126表达(P＜0.05),抑制95D细胞的生长和迁移能力(P＜0.05).Western Blot结果表明细胞中磷酸化AKT水平显著降低(P＜0.05).结论:通过上调miR-126的表达水平可有效抑制95D细胞的体外生长,其作用机制有可能与AKT信号通路的变化有关,但其具体机制仍需后续深入研究.     

小檗碱对三转基因阿尔茨海默病小鼠的学习记忆和海马PSD95突触相关蛋白表达水平的影响

目的 探讨小檗碱(BBR)对三转基因阿尔茨海默病(AD)小鼠的学习记忆及海马组织PSD95突触蛋白表达水平的影响。方法 将30只三转基因(APP/Tau/PS1)AD小鼠按随机数字表法分成3组,即AD对照组、AD+25 mgBBR组、AD+50 mgBBR,每组各10只,后2组以灌胃方式且剂量分别为25 mg·kg^-1·d^-1、50 mg·kg^-1·d^-1,对照组给予等剂量生理盐水连续3个月灌胃处理; 采用Morris水迷宫方法探测各组AD小鼠行为学改变、空间记忆及探索情况; 免疫荧光染色检测各组小鼠海马组织突触后致密蛋白95(PSD95)阳性表达水平; Western blotting(WB)法检测各组三转基因AD小鼠海马脑组织PSD95蛋白、磷酸化蛋白激酶B(p-Akt)和磷酸化雷帕霉素靶蛋白(p-mTOR)表达水平及微管相关蛋白轻链3-Ⅱ(LC3-Ⅱ)自噬水平。结果 AD+25 mgBBR组的逃避潜伏期的学习记忆能力、免疫荧光PSD95表达水平以及PSD995、LC3-Ⅱ、p-Akt、p-mTOR蛋白表达水平与AD对照组比较均有明显差异(P<0.05); AD+50 mgBBR组逃避潜伏期的学习记忆能力、免疫荧光PSD95表达水平以及LC3-Ⅱ、p-Akt、p-mTOR表达水平与AD对照组比较差异均更明显(P<0.05,P<0.01)。结论 应用50 mg小檗碱能较好改善三转基因AD小鼠的学习记忆、空间探索能力,其机制可能是通过增加自噬水平LC3-Ⅱ调控Akt/mTOR信号通路,增加突触蛋白PSD95的表达水平及突触数量,以改善AD相关临床症状。     

TGF‐β2 and TGF‐β3 from cultured β‐amyloid‐treated or 3xTg‐AD‐derived astrocytes may mediate astrocyte–neuron communication

Astrocytes participate in the development and resolution of neuroinflammation in numerous ways, including the release of cytokines and growth factors. Among many, astrocytes release transforming growth factors beta (TGF‐β) TGF‐β1, TGF‐β2 and TGF‐β3. TGF‐β1 is the most studied isoform, while production and release of TGF‐β2 and TGF‐β3 by astrocytes have been poorly characterized. Here, we report that purified cultures of hippocampal astrocytes produce mainly TGF‐β3 followed by TGF‐β2 and TGF‐β1. Furthermore, astrocytes release principally the active form of TGF‐β3 over the other two. Changes in release of TGF‐β were sensitive to the calcineurin (CaN) inhibitor FK506. Starvation had no effect on TGF‐β1 and TGF‐β3 while TGF‐β2 mRNA was significantly up‐regulated in a CaN‐dependent manner. We further investigated production and release of astroglial TGF‐β in Alzheimer's disease‐related conditions. Oligomeric β‐amyloid (Aβ) down‐regulated TGF‐β1, while up‐regulating TGF‐β2 and TGF‐β3, in a CaN‐dependent manner. In cultured hippocampal astrocytes from 3xTg‐AD mice, TGF‐β2 and TGF‐β3, but not TGF‐β1, were up‐regulated, and this was CaN‐independent. In hippocampal tissues from symptomatic 3xTg‐AD mice, TGF‐β2 was up‐regulated with respect to control mice. Finally, treatment with recombinant TGF‐βs showed that TGF‐β2 and TGF‐β3 significantly reduced PSD95 protein in cultured hippocampal neurons, and this effect was paralleled by conditioned media from Aβ‐treated astrocytes or from astrocytes from 3xTg‐AD mice. Taken together, our data suggest that TGF‐β2 and TGF‐β3 are produced by astrocytes in a CaN‐dependent manner and should be investigated further in the context of astrocyte‐mediated neurodegeneration.     

孤独症大鼠前额叶皮质突触相关蛋白的表达

目的 观察孤独症大鼠突触相关蛋白突触素(SYN)、突触后致密蛋白-95(PSD-95)、桥尾蛋白(gephyrin)在前额叶皮质的表达变化,探讨突触相关蛋白在孤独症发病中的作用。方法 采用孕12.5 d(E12.5)一次性腹腔注射丙戊酸钠(600 mg/kg)的孕鼠所产下的子代鼠作为模型组,同样方法注射同等剂量的生理盐水的孕鼠所产下子代鼠作为正常组。通过斜板实验、睁眼实验、三箱实验验证模型是否成功;Western blotting方法对比生后42天两组大鼠突触相关蛋白SYN、PSD-95、gephyrin在前额叶皮质的表达变化。结果 1)成功建立孤独症动物模型:与正常组比较,模型组大鼠生长发育迟缓;社会交往能力异常、对新鲜事物偏好障碍,差异均有统计学意义(P均<0.05);2)Western blotting结果显示:与正常组比较,孤独症大鼠突触相关蛋白SYN、PSD-95表达显著增多(P<0.05),gephyrin蛋白表达显著减少(P<0.05)。结论 孤独症大鼠前额叶皮质突触蛋白表达异常。     

Ocular autofluorescence in diabetes mellitus. A review

Diabetes mellitus is a metabolic disease with a considerable impact on healthcare owing to its increased prevalence and high mortality rate. Structural, morphological, and physiological changes in each of the ocular components have been described in detail. Autofluorescence has been described as a good indicator of metabolic activity. The aim of the present review is to provide an overview of ocular endogenous fluorophores in the cornea, the crystalline lens, and the retinal pigment epithelium, the effects of diabetes mellitus and therefore the potential of autofluorescence assessment for screening and monitoring changes in diabetic patients.     

棘球蚴(包虫)病预防疫苗的研制与应用

棘球蚴(包虫)病的控制是一个长期的过程。20世纪90年代以前,寄生虫病的防治手段主要是药物防治,但近年来,由于寄生虫耐药风险的增加和不理想的控制效果,以及大面积药物的使用对环境的不良影响等,寄生虫病控制的研究方向逐渐向免疫预防转移。本文从棘球绦虫疫苗研制的策略,中间宿主和终末宿主抗虫疫苗的研制,以及所面临的挑战和机遇等做一综述。在我国,EG95已经大面积接种绵羊用于囊性包虫病的控制。但泡球蚴病病原的传播是循环于犬,狐狸和狼等终末宿主和鼠类之间,终末宿主抗虫疫苗研制是泡球蚴病控制研究的方向之一。