To reveal the effects of small molecule drug Cidofovir（CDV） in the transduction of the recombinant adeno-associated virus type 2（rAAV2）. Different doses of CDV were used to intervene rAAV2 vector transduction of different reporter genes（GFP and LacZ） in Hela cells. The effect of CDV on rAAV2 transduction was studied from the aspects of transgene expression, positive cell number and gene number in Hela cells. The experimental results showed that CDV could promote rAAV2 transduction, and the differences in transgene expression, positive cell number and genome number were statistically significant compared with those in the control ground.Furthermore, the time-dosage dependent relationship existed in the transduction efficiency of rAAV2 in Hela cells.

该文探讨了建立生物学特性稳定的人胎盘底蜕膜间充质干细胞(DB-MSCs)库的可行性,旨在为组织工程种子细胞提供更多来源.该研究采用组织块贴壁法从10例足月人胎盘底蜕膜组织中获取间充质干细胞,并采用STR检测细胞是否均来源于母体组织.对分离得到的DB-MSCs采用-196℃低温冻存,并在一定时间复苏培养,用倒置显微镜观察细胞形态,CCK8法检测细胞增殖能力,流式细胞仪分析细胞周期和细胞表面标志物,特定培养基诱导其成脂、成骨、成软骨分化.结果显示,从底蜕膜组织中分离培养扩增获得数目稳定的DB-MSCs.STR分析证明,所得细胞均来源于母体组织,经冻存并复苏后的DB-MSCs形态和表面标志物保持不变,并可稳定扩增8代以上,倍增时间为(2.04±0.25)天,大多数的细胞处于静止期(G0/G1),复苏后的细胞保持了较强的成脂、成骨和成软骨分化能力,在细胞中未发现细菌、真菌、支原体和内毒素等的污染.该研究初步建立了DB-MSCs库,进一步明确了库存细胞的制备流程及其质量评价体系,可为再生医学修复重建组织工程提供较好的种子细胞.

为构建应用型本科院校,提高细胞生物学实验教学水平,以细胞生物学实验课程改革实践为例,探析新建应用型本科高校细胞生物学实验教学存在的问题及改革方法,不断优化教学内容,丰富教学方式与手段,完善教学条件与考核.促进应用型本科高校细胞生物学实验教学示范中心建设,保障研究性教学的实施,提高学生自主学习和独立研究的能力.

White shrimp Litopenaeus vannamei are widely cultured in the world and white spot syndrome virus (WSSV) led to huge economic losses in the shrimp industry every year. In the present study, miRNAs involved in the response of shrimp L. vannamei to WSSV infection were obtained through the Illumina HiSeq 2500 high-throughput next-generation sequencing technique. A total number of 7 known miRNAs and 54 putative novel miRNAs were obtained. Among them, 14 DEMs were identified in the shrimp infected with WSSV. The putative target genes of these DEMs were related to host immune response or signaling pathways, indicating the importance of miRNAs in shrimp against WSSV infection. The results will provide information for further research on shrimp response to virus infection and contribute to the development of new strategies for effective protection against WSSV infections.

为了开发利用绿潮藻类条浒苔中含量丰富的蛋白质,采用木瓜蛋白酶酶解条浒苔蛋白,最佳酶解条件为:料液比1:25、加酶量1250 U/g pro、温度45.7℃、pH 7.2、震荡酶解时间120 min.在该条件下的酶解多肽经超滤分离获得不同分子量区间的多肽,并通过HUVEC、A549、H446、H460等细胞水平初步评价酶解条浒苔制备多肽抗肺癌的细胞生物学效果.结果表明,经上述条件获得的2-6 kD条浒苔多肽处理的HUVEC在抗氧化和小管形成等方面均受到抑制,其处理的NCI-H460、NCI-H446和A549的增殖、细胞周期、迁移也受到不同程度抑制,且能够促进细胞凋亡;2-6 kD浒苔多肽对H446、H460作用效果显著优于对A549的作用.这一细胞水平研究,不仅证实了酶解条浒苔获取新型天然抗肿瘤小肽的可行性,也为条浒苔蛋白质资源高值化利用探索提供了新的途径.

Iron extensively exists in brain tissues, iron is needed for mitochondrial respiration and synthesis of myelin, neurotransmitters. Accumulation of iron in the brain increases with age. Excess iron in the brain is associated with diseases like Alzheimer's, Parkinson's disease, leading to the formation of reactive oxygen species and ER stress. Rabs are molecular switches controlling multiple steps in membrane traffic. Rab8A is founded to be a regulator of glucose and rhodopsin transport recently. In the present study, We found Rab8A is reduced in iron overloaded neural cells, indicating a mechanism of Alzheimer's and Parkinson's disease.

Objective To promote the induction and separation efficiency of bone marrow-derived dendritic cells (BMDCs) in vitro through optimizing the inducing and isolating process by multiple cytokines. Methods The factors to be optimized in single factor tests included recombinant mouse granulocyte macrophage-colony stimulating factor (rmGM-CSF), recombinant mouse interleukine 4 (rmIL-4), lipopolysaccharide (LPS), recombinant mouse tumor necrosis factor α (rmTNF-α) and inducing time. The numbers of immature dendritic cells (imDCs) and mature dendritic cells (mDCs) were investigated as the indicators. Box-Behnken experimental design-response surface methodology was used to analyze and verify the data. Morphological changes were observed using the inverted microscopy and the transmission electron microscopy. Surface molecules including CD11c and CD86 were detected using the flow cytometry. Results The optimum inducing conditions for imDCs were obtained as follows: rmGM-CSF was 46 ng/mL, rmIL-4 was 24 ng/mL, inducing time was 6 days, and the number of imDCs was (4.58±0.28)×106 cells, and the relative deviation was 4.00%. The optimum inducing conditions for mDCs were as follows: LPS was 1.4 μg/mL, rmTNF-α was 30 ng/mL, inducing time was 1 day, and the number of mDCs was (4.21±0.15)×106 cells, and the relative deviation was 3.80%. Sufficient typical imDCs and mDCs were obtained within 5-7 days of induction in vitro. Also, flow cytometry showed that the amplified imDCs had a high expression of CD11c (68.62%±2.3%) and a low expression of CD86 (37.95%±1.8%), and the mDCs had high expressions of both CD86 (90.34%±1.4%) and CD11c (82.05%±1.6%). Conclusion The combination of single factor tests and Box-Behnken design -response surface methodology could optimize the inducing and isolating method for DCs in vitro by multiple cytokines rapidly and efficiently, which provided basic experiment materials for further studies.

This study was designed to evaluate the anti-inflammatory effect of recombinant human kallistatin (Kal) on ulcerative colitis (UC) in the mouse model. Acute colitis was induced by administration of 4% dextran sodium suffate (DSS) to KM mice for 7 days. The mice were then randomized into 5 groups: model control, Kal 0.2 mg·kg(-1)·d(-1), 1.0 mg·kg(-1)·d(-1) and 2.0 mg·kg-1·d(-1) group, salazosulfapyridine (SASP) group. Ten age-matched normal KM mouse were administered with saline in the normal control. The weight, colon length, inflammation factor (MPO/SOD/MDA) and TNF-α/IL-10 levels among the five groups of mice were determined. The results showed that histological index score and MPO/MDA/TNF-α levels of high-dose Kal treatment group and SASP group were significantly lower compared with the model group (P < 0.01), but the weight, colon length, IL-10 level and SOD activity were significant higher than the model group (P < 0.01), approaching the normal group. These parameters showed that Kal can significantly relieve the UC state in a dose-dependent manner. This study demonstrates that Kal significantly remits UC in mice, and participates in the regulation of inflammatory cytokines TNF-α/IL-10 levels and has some antioxidant activity.

Haemophilia A is an X-linked recessive monogenic hereditary bleeding disorder caused by a deficiency or functional defect in coagulation factor VIII (FVIII). Typically, only 30% haemophilia A patients are treated with FVIII-specific products successfully. Therefore, other promising clotting factors and FVIII-bypassing factors exhibiting sufficient FVIII-independent activity, low immunogenicity and prolonged half-life are needed to conquer this malady. Here, we will systematically review the current status of the diverse FVIII-bypassing factors for the treatment of FVIII-insensitive haemophilia A patients.

Human tissue kallikrein-binding protein (Kallistatin, KAL), a secretory protein that participates in the regulation of multiple signaling pathways by binding to the extracellular receptor, however, at present has not been reported about the intracellular activity, and whether it has the similar biological activity with extracellular activity. Here we constructed no signal peptide KAL (NSK) into the adeno-associated virus vector to explore the intracellular activity of KAL. Both the endothelial cell and lung cancer cells could express KAL, but not secreted after rAAV2-NSK transfection. The proliferation and migration of human umbilical vein endothelial cells (HUVECs) were inhibited, but the apoptosis rate was not affected. The proliferation rates, mobility and tubule formation of all the three tested lung cancer cells, such as NCI-H446, NCI-H460 and A549, were inhibited to different extents. This cellular study not only confirmed the intracellular activity, but also suggested it may serve as a kind of "balance factor" in multi-targeted controlling, which may provide a new train of thoughts to explain the regulatory contradiction in PI3K-Akt signaling pathways by KAL.