Protective effect of rAAV2-NTF2 on blood-retinal barrier damage in diabetic rats

AIM: To explore the effect of rAAV2-NTF2 on the blood-retinal barrier damage in early stage diabetic retinopathy. METHODS: Retinal vasculature was observed by Evans blue. NTF2 gene was cloned into adeno-associated virus vector pSNAV. The recombinant pSNAV-NTF2 was transfected into BHK cells. After purification, high-titer rAAV2-NTF2. rAAV2-NTF2 at dose of 4 μL (titer 1.0×1012) were injected into left eyes and rAAV2-EGFP at the same does and titer were injected into the right eyes of 36 rats. After 1 month, diabetes mellitus were induced by STZ. One month after onset of diabetes, EB at a dose of 45 mg/kg was intravenously injected into the rats. Two hours later, both eyes were enucleated immediately after perfusion of 1% PFA-citric acid buffer solution. The retinas were then dissected away and placed in formamide to extract the dye. The concentration of dye was measured by spectrophotometer. RESULTS: Evans blue was contained in normal retinal vessels without leakage, with a very low level of background fluorescence. The vessels staining of diabetic rats’ retina showed increasing fluorescence, indicating the retina-blood barrier damage. Dye concentration, representing the degree of the BRB injury, was higher in retina of 1 month diabetic rats than that in normal rats (4.67 times, P<0.01), indicating retina-blood barrier break-down. Diabetic rats with rAAV2-NTF2 intravitreal injection showed decreased BRB breakdown (P<0.05). CONCLUSION: High expression of NTF-2 decreases the BRB dysfunction in DM rats.     

Asymmetric dimethylarginine protects PC12 cells against glutamate-induced excitotoxic damage

AIM: To investigate the effects of asymmetric dimethylaoyoinine (ADMA) on glutamate-induced PC12 cell damage and its mechanisms. METHODS: PC12 cells were treated with different concentrations of glutamate as an in vitro excitotoxic trauma model. The cell viability was measured by MTT assay. Glutamate cytotoxicity was evaluated by lactate dehydrogenase (LDH) release assay. Intercellular reactive oxygen species (ROS) was detected by dihydrorhodamine123 (DHR) staining and flow cytometric (FCM) analysis. Nitric oxide synthase (NOS) activity and nitric oxide (NO) production were detected by using commercial kits with a spectrophotometer. RESULTS: Glutamate at concentrations of 1 mmol/L to 6 mmol/L dose-dependently decreased PC12 cell viability. Pretreatment 30 min with ADMA prior to administration of glutamate significantly attenuated the inhibition of cell viability, LDH release and ROS accumulation induced by glutamate. Pretreatment with ADMA significantly inhibited the increases in NOS activity and NO production caused by glutamate. CONCLUSION: ADMA obviously protects PC12 cells against glutamate-induced excitotoxicity by inhibiting NOS activity, overproduction of NO and accumulation of intracellular ROS.     

富士苹果AFLP体系的优化及其在鉴定早熟芽变中的应用

 以‘长富2号’苹果为材料,采用优化的AFLP技术对其早熟芽变与母株进行初步研究,筛选了64对引物,其中43对引物扩增结果理想,共产生了2,700条带。有4对引物在芽变与母株之间产生了7条差异片段,主要集中在350~800 bp之间。研究表明早熟芽变是由于遗传物质改变造成的,同时也说明利用该体系对苹果芽变进行分析和鉴定获得成功。     

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.     

华蔓茶藨子扦插繁殖技术研究

研究了华蔓茶藨子枝条不同部位、IBA不同浓度处理对扦插生根的影响.结果表明:插穗不同部位对愈伤组织出现期无影响,均为5 d,在生根时间上存在差异,上部16 d生根,比中部和基部提前2 d生根;对根质量的影响差异显著,上部最好,中部次之,下部最差;NAA和IBA对插穗生根均有明显促进作用,且IBA优于NAA,100 mg/L mA生根效果最好,生根率和生根质量均达到最高.     

干旱胁迫对果树开花的诱导及对生理生化性状的影响

干旱胁迫是许多热带、亚热带果树花芽诱导的有效措施,现就干旱胁迫对果树开花诱导的效果以及对果树生理生化过程的影响进行讨论.认为干旱胁迫明显降低果树的营养生长,影响果树体内碳水化合物以及含氮化合物的含量水平和相对比例,明显调节果树体内内源激素含量状况,并有效促进部分果树的开花.同时对干旱诱导的时间、强度以及今后控水促花研究中尚需解决的问题进行了讨论.     

西瓜、甜瓜果实品质及调控研究进展

西瓜甜瓜深受消费者喜爱,但是近年来品质呈现下降的趋势.西瓜甜瓜的品质包括外观品质、营养品质和风味品质,而品质的形成又受品种、土壤环境、肥料供应、水分供应、栽培方式、生长调节剂以及采收与贮运等因素的影响,所以要针对以上因素,从改进育种手段和栽培技术2大方面来改良和提高品质.     

华西雨屏区5种坡地利用方式产流产沙与养分流失特征

选择5种坡地利用方式,分别是裸地、农耕地(玉米)、荒草地、巨桉林地Ⅰ与巨桉林地Ⅱ,建立人工径流小区,对2006年6～11月各小区产流产沙及养分流失进行对比研究.结果表明:(1)5种坡地利用方式产流产沙差异显著(P＜0.05),径流深依次是裸地(250.9 mm)＞农耕地(232.9 mm)＞荒草地(175.4 mm)＞巨桉林地Ⅰ(170.5 mm)＞巨桉林地Ⅱ(141.3 mm);产沙量大小依次为:农耕地(313.63 t/km2)＞裸地(176.80 t/km2)＞荒草地(94.45 t/km2)＞巨桉林地Ⅰ(90.58 t/km2)＞巨桉林地Ⅱ(36.37 t/km2).(2)5种坡地利用方式养分流失差异显著(P＜0.05),其流失量大小顺序均为钾素＞磷素＞氮素.其中,氮素与磷素流失以水溶态为主,平均为96.16%与69.45%;钾素以颗粒态(与泥沙结合态)为主,平均为74.93%.(3)不同坡地产流产沙主要影响因素不同,裸地与农耕地受降雨影响较大,而荒草地与巨桉林地受人为干扰因素较大.(4)调整农事活动、改善经营措施与减少人为干扰对防止坡地产流产沙以及养分流失具有重要作用.     

植物篱种植模式及其在我国的研究和应用

植物篱种植模式作为一种坡地保护性耕作和持续利用技术,自20世纪70年代以来在热带地区研究和应用较为广泛,本文概述了我国近10余年来的研究和应用进展。这一模式在20世纪80年代中期进行了短暂的试验,20世纪90年代以来在金沙江干旱河谷地区和三峡地区结合当地实际需求和植物篱种植模式的特点,对植物篱种植模式进行了研究,使之适合我国山区和坡耕地的应用;对植物篱的生态效益、社会效益和经济效益均进行了较深入细致的试验、研究,并进行较大面积的示范和推广应用,同时引起了国内很多研究机构和应用部门的关注,尽管在1997年以后其它区域也随之进行了一些类似的试验和研究,但国内目前只有金沙江干旱河谷地区和三峡地区在研究和应用方面较系统和深入。最后还对国内在植物篱认识上的一些问题,以及需要进一步深入开展的工作进行了讨论。     

华西雨屏区人工竹林凋落物及表层土壤的水源涵养功能研究

以华西雨屏区退耕还林后形成的人工竹林(慈竹林、苦竹林、撑绿杂交竹林、苦竹 光皮桦混交林)为对象,对其凋落物及表层土壤的水源涵养功能进行了初步研究。结果表明:几种竹林凋落物蓄积量的大小顺序是:撑绿杂交竹林>混交林>苦竹林>慈竹林,且都大于光皮桦林。叶凋落物持水量的大小关系为:慈竹林>混交林>苦竹林>杂交竹林,光皮桦林的持水量介于慈竹林和混交林之间。几种叶凋落物持水量与浸泡时间的关系符合指数函数的模型、持水速率与浸泡时间的关系符合幂函数的模型。不同林地的表层土壤容重在1.27~1.34 g/cm3之间,总孔隙度48.08%~55.96%,不同林地表层土壤的水分物理性质差异不大。