菠萝锌指蛋白基因AcRCHY1的克隆与表达分析

 根据植物C3HC4型兼CHY型锌指蛋白的功能保守区设计简并引物, 通过RT2PCR结合RACE 方法从菠萝(Ananas comosus L. Merr) 幼苗中克隆获得了一个新的菠萝锌指蛋白基因cDNA全长, 将其命名为AcRCHY1。该基因cDNA全长1 261 bp, 开放阅读框ORF为918 bp, 推测其编码一个含有306个氨基酸残基的多肽。AcRCHY1蛋白具有保守的C3HC4 (RING finger) 和CHY两个锌指结构域, 与其它植物锌指蛋白的同源性高达81%～91%。半定量RT-PCR分析表明, AcRCHY1 在菠萝中呈组成性表达, 在子房、花瓣和小花中的表达量明显高于根和叶; 低温、高盐、干旱和ABA等非生物胁迫处理后, AcRCHY1在叶片中的表达明显增强。因此, AcRCHY1蛋白可能与花器官生长发育的调控有关, 而且可能作为一个转录调控因子在菠萝响应低温、高盐和渗透胁迫过程中参与了依赖ABA的信号转导途径。     

Identification of serum differential proteins in colorectal adenoma and early malignantly transformed adenoma by proteomics

AIM: To investigate the associated proteins and sensitive biomarkers for early diagnosis of colorectal adenocarcinoma by comparing the results of differential proteomic analysis between colorectal adenoma and early malignantly transformed adenoma. METHODS: Two-dimensional gel electrophoresis was used to define patterns of protein expressions of colorectal adenoma and early malignantly transformed adenoma. Proteins expressed differentially among groups were detected, cut out and analyzed by MALDI-TOF/TOF mass spectrometry. RESULTS: Two-dimensional protein maps of colorectal adenoma and early malignantly transformed adenoma were analyzed with gel-analysis software, an average of 1 672 spots in adenoma, 1 732 in early malignantly transformed adenoma were observed. 28 spots of a 1.5-fold change were found, including 15 proteins down-regulated and 13 up-regulated in early malignantly transformed adenoma, in which 23 proteins were identified by mass spectrometry, the rate of identification was 82.14%. 13 differential proteins were attained, 8 were up-regulated and 5 were down-regulated, which was classified to 6 categories, including protease inhibitor, complement, immunoglobulin, keratoproteins, signal transduction protein and function-unknown proteins. CONCLUSION: The changes of serum proteins in early malignantly transformed adenoma from adenoma can be identified by proteomic technology. Proteins detected in the study may provide new biomarkers correlated with biological behavior of colorectal adenocarcinoma.     

银杏叶对辣椒植株发酵液自毒作用的化感调控

采用1/2霍格兰（Hoagland）营养液水培法,研究辣椒植株发酵液的自毒作用及银杏叶发酵液和 再腐解液对其的调控作用。结果表明,辣椒植株发酵液使辣椒受到自毒作用,根系活力下降,相对电导 率增大。银杏叶发酵液加重了辣椒植株发酵液的自毒作用,并且与其浓度成正相关。银杏叶再腐解液缓 解了辣椒的自毒作用,缓解效果与再腐解液的浓度有关。     

新疆野苹果群体遗传结构和遗传多样性的SRAP分析

 采用SRAP标记,对中国新疆野苹果4个种下居群的群体遗传结构和遗传多样性进行了研究。结果表明：10对SRAP引物总共扩增了209条带,其中206条是多态性带（98.56%）。巩留群体、新源群体、霍城群体和裕民群体分别扩增了180、169、178和165条多态性带,巩留群体的随机交配杂合度（hs = 0.3037 ± 0.0058）最高,其次为霍城群体。UPGMA聚类分析和群体间遗传分化系数显示,巩留群体和新源群体之间,以及霍城群体和裕民群体之间,遗传关系最近,群体间遗传变异最低。新疆野苹果群体内遗传变异高于群体间,占总变异的87.9%,主坐标轴分析显示4个群体是相对独立,其中巩留群体和新源群体,霍城群体和裕民群体之间,有较高的基因交流。所有参数分析表明,巩留群体遗传多样性最丰富,故在制定新疆野苹果原地和异地种质保护计划时应优先考虑巩留群体。     

Abnormalities of miR-21 expression in human breast cancer is associated with distinctive pathologic features and shortened postoperative survival

AIM: To investigate the potential relevance of miR-21 expression level to clinicopathological characteristics and patient survival. METHODS: 113 BRCA cases with more then 5 years fallow-up data were selected. Total RNA from formalin-fixed paraffin-embedded (FFPE) tissues of 113 breast cancer (BRCA) and normal adjacent tissues (NATs) were isolated for miR-21 quantitative analysis by real-time RT-PCR. RESULTS: The miR-21 expression levels in BRCA were significantly higher than those in NATs (P<0.01) with average up-regulated level of 1.74 ± 0.48. Interestingly, high level expression of miR-21 was significantly correlated with advanced clinical stage (P<0.01), lymph node metastasis (P<0.01), and shorter survival of the patients ［hazard ratio (HR)=5.476, P<0.01］. Multivariate Cox regression analysis revealed that miR-21 was one of independent prognostic impacts (HR=4.133, P<0.01) on BRCA. CONCLUSION: Over-expression of miR-21 is associated with poor prognosis of BRCA and may serve as an independent prognostic marker for BRCA.     

Relationship between endothelial dysfunction and renin-angiotensin-aldosterone system under the condition of fatigue stress and effect of herbs to dredge collaterals

AIM: To investigate the mechanisms and relationship among endothelial dysfunction, renin-angiotensin-aldosterone system (RAAS) and family of interleukins under the condition of excessive fatigue, by using the rats with fatigue stress. METHODS: Healthy male Wistar rats were randomly divided into control group, homocysteine (HCY) group, fatigue stress group, renshen group, shuangshen group and tongxinluo group. Radioimmunoassay was carried out to detect plasma renin activity (PRA), angiotensin II (AngⅡ), aldosterone (ALD), endothelin (ET), thromboxane-2 (TXA2), prostaglandin I2 (PGI2) in plasma and interleukin-1β, 2, 6, 10 (IL -1β, IL -2, IL -6) in sera. ELISA was used to detect NE and IL -10. The content of nitric oxide (NO) in sera was also detected. The bioinformatical analysis was used to determine the relationship between RAAS and endothelial dysfunction. RESULTS: Compared with control group, the levels of ET-1 and TXA2 in fatigue stress group were significantly increased (P<0.01, P<0.05), but the content of PGI2 and NO was significantly decreased (P<0.01, P<0.01). Compared with control group, the renin activity in plasma of animals in fatigue stress group was significantly decreased (P<0.01), the AngⅡ, IL -1β, IL -6 level was significantly increased compared with the control group (P<0.01, P<0.01), and was significantly increased compared with the HCY group (P<0.05, P<0.01, P<0.01). The NE level showed the tendency of decrease in different degree. After the intervention of three kinds of herbs to dredge collaterals, the ET-1, AngⅡ, IL -1β, IL-6 level in plasma was decreased in different degree (P<0.05, P<0.01, P<0.01), and at the same time, the contents of NO and NE level were significantly increased (P<0.05). The ALD level in tongxinluo groups was apparently higher than that in control group and the fatigue stress group (P<0.05). The bioinformatics analysis showed that Ang II and ET, IL-1; PGI2 and ALD; NO and ALD composed of three subsystems and interrelated according to the principle of optimality of complex system, and gradual change regulation was also observed in fatigue stress group. However, in control group, HCY group and tongxinluo group, the same interrelation among subsystems was not existed. CONCLUSION: In a state of long-term excessive fatigue, vascular endothelial dysfunction may be induced, and is related with renin-angiotensin-aldosterone system imbalance and serious turbulence of the autonomic dysfunction. Herbs to dredge collaterals could improve it significantly. The results suggest that bearing excessive fatigue and pressure in long-term may be the potential risk factors to induce vascular endothelial dysfunction and further result in cardio-cerebro vascular diseases, Tongluo therapy may be one of the useful ways to prevent such diseases.     

Effects of diabetes on the development of heart failure in streptozotocin-induced diabetic rat model with acute myocardial infarction

AIM: To evaluate the time course and effect of diabetes on the development of heart failure (HF) in poorly controlled streptozotocin (STZ) -induced diabetic rat model for 70 d with acute myocardial infarction (AMI) in vivo. METHODS: All SD rats were randomized into four groups. Diabetes were induced by a single intraperitoneal injection of STZ (65 mg/kg), and 70 d later after the induction, AMI models were made with the ligation of left anterior descending coronary artery. The time course of diabetic effects on the development of heart failure in rats before and after AMI was observed. The survival rate of the rats and ultrastructure change of myocardium, the hemodynamics, the extent of the myocardial fibrosis, and the cardiac hypertrophy were also determined. RESULTS: After the ligation of left anterior descending coronary artery, the diabetic rats showed worse LV function and accelerated left ventricular (LV) remodeling compared with the non-diabetic ones. Myocardial fibrosis in both diabetic and non-diabetic rats subjected to AMI was similar in the early phase, while it was quite different after 1 month. CONCLUSION: Heart failure progression is accelerated in diabetic rat with AMI.     

洋葱细胞质雄性不育系选育研究进展

 介绍了国内外洋葱细胞质雄性不育（CMS）系的选育及利用概况;对洋葱细胞质雄性不育机理和相关分子标记进行了综述;在此基础上对洋葱雄性不育的研究提出了几点建议。     

Jagged1 knocking down in endothelial cells accelerates PDGF induced proliferation and migration of vascular smooth muscle cells in rats

AIM: To investigate the effect of Jagged1 expression in endothelial cells (EC) on platelet derived growth factor (PDGF) induced proliferation and migration of vascular smooth muscle cells (VSMC) in rat.METHODS: Rat aorta EC was inoculated in the lower chamber and VSMC were in the upper chamber of the cell coculture system. Three groups were divided: control, sicontrol and siJagged1. The EC Jagged1 protein expression was assayed by Western blotting to evaluate small RNA interfering (RNAi) efficiency. After the cells were cocultured with PDGF for 24 h, the proliferation and migration of VSMC were respectively evaluated by ［3H］-TdR incorporation and migrating cells counting. Protein expression of α-SM-actin in VSMC was assayed by Western blotting. RESULTS: The Jagged1 protein expression in EC was significantly lower in siJagged1 group than that in control group (0.26±0.02 vs 0.67±0.02, P<0.05), and no statistic significance was observed between control and sicontrol groups. The VSMC ［3H］-TdR incorporation and migration were higher in PDGF +siJagged1 group than those in PDGF group {［3H］-TdR incorporation (23 074±2 702) counts·min-1·well-1 vs (16 442±1 803)counts·min-1·well-1, n=5, P<0.05; migration (27±4) cells/field vs (15±3)cells/field, n=5, P<0.05}. The α-SM-actin protein in VSMC was lower in PDGF + siJagged1 group than that in PDGF group (0.25±0.06 vs 0.49±0.04, n=3, P<0.05).CONCLUSION: Jagged1 knock down in rat EC accelerates PDGF induced proliferation and migration of VSMC. These results suggest that Jagged1 expression in EC plays an important role in maintaining VSMC contract phenotype and inhibiting VSMC overgrowth after arterial injury.