Generation of insulin-producing cells from PDX1 and NKX6.1 gene-modified bone marrow mesenchymal stem cells

AIM: To differentiate bone marrow mesenchymal stem cells (BMSCs) into functional insulin-producing cells to produce sufficient pancreatic islet cells for transplantation. METHODS: Recombinant adenovirus vectors carrying PDX1 and NKX6.1 genes were constructed and the bone marrow mesenchymal stem cells were infected by the recombinant adenovirus combined with several cytokines for differentiation. The expressions of PDX1, NKX6.1 and insulin and C-peptide in the differentiated bone marrow mesenchymal stem cells were detected by RT-PCR and Western blotting. After the differentiated bone marrow mesenchymal stem cells were transplanted into subrenal capsule of diabetic mice, cell morphology of the grafts as well as their secretion of insulin and C-peptide were detected. Besides, regulating capacities of grafts on the blood glucose level of the diabetic mice were also detected. RESULTS: BMSCs induced by recombinant adenovirus (pAdxsi-CMV-PDX1/CMV-NKX6.1) and several cytokines showed positive dithizone staining and the expressions of β-cell related molecule such as insulin and glucose transporter 2 were detected by RT-PCR, which showed a sustaining and stable expression. The similar results were showed by Western blotting, immunohistochemical staining and indirect immunofluorescence. The insulin secretions in the cells stimulated with glucose at concentrations of 5.5 and 25 mmol/L in the experimental group were (1 240.4±109.3) mU/L and (3 539.8±245.1) mU/L, respectively, and were significantly higher than those in control group. Moreover, transplantation of the cells to STZ mice in treatment group made serum glucose recover to normal level. CONCLUSION: PDX1 and NKX6.1 gene-modified bone marrow mesenchymal stem cells differentiate into insulin-producing cells in vitro. When these cells transplanted into STZ induced diabetic mice, their serum glucose could return to the normal level and they could live well. Thus this is a promising method for diabetes treatment.     

Inhibitory effect of T-bet gene transfer on airway inflammation in a established murine allergic asthmatic model

AIM: To investigate the effect of T-bet plasmid gene transfer to airway on allergen induced airway inflammation in a murine asthmatic model. METHODS: A mouse asthma model was established by sensitization with ovalbumin (OVA). Forty C57BL/6 mice were divided into 4 groups (10 mice in each group): the normal control group (group A), the asthmatic model group (group B), the pcDNA3 plasmid group (group C), and the pcDNA3-T-bet group (group D). The animals in group B, C and D were sensitized and challenged with OVA. The animals in group A were applied with normal saline. pcDNA3 plasmid at dose of 50 μg was intranasally administered at 24 h before intranasal challenges to the mice in group C, and the 50 μg pcDNA3-T-bet plasmid for the mice in group D. Bronchial alveolar lavage fluid (BALF) was collected and lung tissues were resected at 48 h after OVA challenge for later assay. RESULTS: After administration with pcDNA3-T-bet plasmid, high level of T-bet expression at 48 h was detected in the lung tissue by Western blotting. In pcDNA3-T-bet treated asthmatic models, histological evaluation revealed the significant suppression of eosinophil peribronchial and perivascular infiltration, and reduction of epithelial damage. The numbers of eosinophils, neutrophils and lymphocytes in BALF from pcDNA3-T-bet treated mice were significantly reduced compared to those in asthmatic control group (P<0.05). The level of IL-4 in BALF was significantly decreased in pcDNA3-T-bet group compared to that in asthmatic control group (P<0.05), while the level of IFN-γ in BALF was significantly increased in pcDNA3-T-bet group. No significant change of inflammation cells and cytokines in pcDNA3 plasmid group and asthmatic control group was observed (P>0.05). CONCLUSION: Intranasal pcDNA3-T-bet plasmid transfer inhibits asthmatic airway inflammation in the murine asthmatic model, suggesting a new therapeutic strategy for allergic asthma.     

Effects of apolipoprotein (a) on vascular smooth muscle cells proliferation and cell signal transduction pathway

AIM: To investigate the effect and the mechanism of apolipoprotein (a) ［apo (a)］ on proliferation of vascular smooth muscle cells (VSMCs). METHODS: All VSMCs used in experiments were serial subcultured from primary cells and were identified by immunohistochemistry staining of α-actin. Cell growth assay was observed as cell counting and MTT assay. Western blotting was also employed to detect the related mechanism. RESULTS: All cells used in experiments were confirmed as VSMCs. Although apo (a) enhanced VSMCs proliferation, this effect was attenuated by anti-integrin αⅤβ3, LM609. Use these reagents alone had no effect on VSMCs growth. The results of Western blotting demonstrated that focal adhesion kinase (FAK) was activated by apo (a) and the expression of total or phosphorylated transforming growth factor β1 (TGF-β1) was also decreased. However, these effects described above were all blocked by LM609. CONCLUSION: Apolipoprotein (a) enhances VSMCs proliferation and this effect is mediated by integrin αⅤβ3, which activates FAK and attenuates TGF-β1 and phospho-TGF-β1 expression.     

Impact of serial activation of embryonic pax2 and WT1 genes on transdifferentiation of renal tubular epithelial cells

AIM: To investigate the mechanisms of transdifferentiation of renal tubular epithelial cells by the observation of serial activation of embryonic pax2 and WT1 genes in chronic renal failure model of 5/6 nephrectomized injury. METHODS: Embryo of mice, cultured renal tubule cells and chronic renal failure rat model of 5/6 nephrectomized injury were established. The expressions of paired Box gene (pax2) and Willm’s tumor gene (WT1), as well as α-smooth muscle actin (α-SMA), a phenotype of mesenchymal cells, were detected by RT-PCR and immunohistochemistry techniques. RESULTS: (1) Expressions of pax2 and WT1 mRNA began at 11.5 d and 14 d of embryo and increased gradually, and expression in trace quantity at 2 weeks after birth. Immunohistochemistry analysis revealed that WT1 only expressed in the glomerular podocytes and expressions of pax2 and WT1 in adult renal tubular cells were negative. (2) Deno-expression of pax2 and WT1 in some tubular cells appeared at week 2 and peaked at week 4 in 5/6 nephrectomized rats, both showing a same trend of expression. However, pax2 showed another peak at week 10 afterwards. (3) Re-expressions of pax2 and WT1 in TEC at 0.5 and 24 h after treated with IL-1α (10 μg/L) or AngII (10-9 mol/L) were observed respectively, followed by upregulation of α-SMA expression, and mesenchymal cells characters were shown. The effects were inhibited by Ang II receptor 2 antagonist and WT1 antibody, respectively. CONCLUSION: Adult renal tubule cells acquire re-activation of embryonic pax2 and WT1 genes and phenotypes of mesenchymes when challenge with certain injuries. Deno-expression of pax2 and WT1 genes closely relates with high concentration of bioactive facors, such as AngII and IL-1, which are involved in the mechanisms of renal tubule cells transdifferentiation.     

硬肉大果型番茄新品种北研1号的选育

北研1号是以05B-91为母本,以05B-93为父本配制而成的硬肉大果型番茄一代杂种。无限生长类型,中早熟,生育期 106 d（天）左右。成熟果红色,扁圆形,无绿肩,可溶性固形物含量4.8 %,VC 114.4 mg·kg^-1,可溶性总糖2.7 %。抗叶 霉病、ToMV,果实硬度0.84 kg·cm^-2,耐贮运,平均单果质量192.7 g,每667 m²产量6 000 kg左右,全国各地保护地、露 地均可栽培。     

冀西北坝上地区几种蚜虫优势种对菜豆的侵害性研究

采用寄主转换方法研究了冀西北坝上地区几种蚜虫优势种对菜豆的适应性。结果表明,桃蚜、瓜蚜（棉蚜）和菊姬长管蚜转接到露地菜豆 的花、嫩尖、嫩叶等部位上仅能存活6~8 d,不能在菜豆上建立种群。室内寄主转换试验表明,取食菜豆嫩荚的蚜虫存活率明显低于取食茎部嫩尖的 蚜虫存活率。     

不同来源菠菜品种营养品质分析与评价

为评价不同来源菠菜品种的营养品质,筛选出营养物质含量丰富,硝酸盐、单宁和草酸含量低的品种,测定了34份 菠菜材料的水分、VC、可溶性糖、粗纤维、草酸、单宁和硝酸盐7个品质指标的含量,并用模糊数学的隶属函数法对其营养品 质进行了综合分析。结果表明：不同菠菜品种在品质方面有较大的差异,34份菠菜材料中,用隶属函数法分析综合营养品质 最好的是欧洲的胜先锋、日本的トライ和アソナ,但在所有品种中,国内的京菠1号VC含量最高,达601.0 mg·kg^-1(FW),且 硝酸盐含量最低。     

广西唇柱苣苔属植物及其园林应用

介绍了广西唇柱苣苔属植物资源的基本特点、观花性状和园林应用,同时对广西唇柱苣苔属植物的保护和开发利用提出了建议.其中,广西唇柱苣苔属植物的6个基本特点是:①唇柱苣苔属观赏植物资源丰富;②观花性状显著;③特有现象突出;④抗逆性特点强;⑤同一种类不同居群其形态变化很大;⑥分布不均性.     

新型保鲜剂处理茂谷柑采后的常温保藏研究

利用不同浓度的新型保鲜剂蔗糖基聚合物和施保克对采后茂谷柑进行涂膜处理后进行常温贮藏,对几项主要生理指标进行检测.结果表明:蔗糖基聚合物涂膜可以减缓Vc、可溶性固形物(TSS)、总糖和可滴定酸等营养物质的损耗,保持较高的糖酸比和固酸比,降低丙二醛(MDA)含量和淀粉酶活性.在延缓果实组织的衰老,延长贮藏寿命方面有较好的效果.施保克也有较好的保鲜效果.