银杏锰型超氧化物歧化酶GbMnSOD基因的克隆与表达

利用RACE技术首次从银杏中克隆到锰型超氧化物歧化酶基因(GbMnSOD ) 的cDNA全长。GbMnSOD的cDNA全长965 bp (GenBank accession number: EF633506) 。生物信息学分析GbMnSOD cDNA序列含有一个681 bp最大读码框, 编码一个226氨基酸多肽链, 通过软件预测分子量为2515 kD, 等电点为8197。三维结构预测结果显示, GbMnSOD 含有12个α螺旋和3 个β折叠构成一个篮子状的活性中心。GbMnSOD氨基酸序列与其它植物MnSOD具有很高的相似性。进化树分析结果表明GbMnSOD和其他物种的MnSOD源自于相同的祖先。Southern杂交显示, GbMnSOD属于一个小的多基因家族。Northern 杂交表明GbMnSOD在银杏的根、茎、叶和果中都有表达, 在叶中的表达量最高, GbMnSOD 的转录受到ABA、IAA、蔗糖、甘露醇、NaCl 和低温的诱导。     

Role of GPR40 mediates the effects of FFAs on lipoapoptosis in mouse β-cell line NIT-1

AIM: To evaluate the role of G protein-coupled receptor 40 (GPR40) mediates the effects of free fatty acids (FFAs) on lipoapoptosis in mouse β-cell line NIT-1 and the mechanisms involved in this process.METHODS: NIT-1 cells were supplemented with palmitate (500 μmol/L) or oleate (500 μmol/L) for 48 h, then apoptosis of the cells was determined by the methods of Hoechst 33342, TUNEL and flow cytometry (Annexin V/PI). The small interfering RNA technique was used to inhibit the expression of GPR40 in NIT-1 cell. The mock, control siRNA and GPR40 siRNA transfected cells were either supplemented with palmitate (500 μmol/L) or co-incubated with palmitate and oleate (500 μmol/L for each) for 48 h. The percentages of apoptotic cells were quantified. The expression of p-c-Jun, Bcl-2 and Bax were detected by Western blotting.RESULTS: Palmitate induced β cell lipoapoptosis, whereas oleate inhibited NIT-1 cells from palmitate-induced lipoapoptosis. No significant difference of the percentages of apoptotic cells was indicated among the mock, control siRNA and GPR40 siRNA transfected cells treated with palmitate (P>0.05). However, after co-incubated with palmitate and oleate (500 μmol/L for each) for 48 h, the percentage of apoptotic cells in GPR40 siRNA transfected cells was greater than that in mock (P<0.05), while the expression of p-c-Jun was decreased. The expressions of Bcl-2 and Bax were not affected.CONCLUSION: Palmitate induced β cell lipoapoptosis might not be mediated through GPR40, whereas oleate inhibits NIT-1 cells from palmitate-induced lipoapoptosis, which is mediated at least in part through GPR40, the change of c-Jun expression may play an role in this process, suggesting that GPR40 might be implicated in the control of β cell mass plasticity and GPR40 probably provides a link between obesity and type 2 diabetes.     

In vitro differentiation of exocrine pancreatic cells from mouse embryonic stem cells

AIM: To explore the effects of sodium butyrate, activin A and dexamthasone on inducing mouse embryonic stem (ES) cells to differentiate into exocrine pancreatic cells in vitro. METHODS: E14 mouse ES cells were cultured in suspension to form embryonic bodies (EBs). The EBs were cultured with differentiating medium containing different concentrations of sodium butyrate, and the spontaneously differentiated ES cells were used as control. Exocrine pancreatic genes such as amylase, chymotrypsinogen, elastase 1, elastase 2 and carboxypeptidase were detected by RT-PCR at different time points to determine the optimal concentration and exposure time of sodium butyrate. Furthermore, activin A or dexamthasone was also used to explore the effects on exocrine differentiation. After that, the combination of sodium butyrate, activin A and dexamthasone was used to promote the differentiation of exocrine pancreatic cells from ES cells. During the differentiation course, the gene expressions of amylase, chymotrypsinogen, elastase 1, elastase 2 and carboxypeptidase were detected by RT-PCR. Morphological changes were investigated by phase contrast microscopy. Amylase expression was examined by immunofluorescence staining. RESULTS: Exocrine pancreatic gene expressions such as amylase, chymotrypsinogen, elastase 1, elastase 2 and carboxypeptidase were detected in spontaneously differentiated EBs. A relatively lower concentration of sodium butyrate with a shorter exposure time significantly promoted those above gene expressions as compared to that of spontaneously differentiated EBs. Activin A and dexamethasone induced upregulation of exocrine gene expression. The combination of activin A, sodium butyrate and dexamethasone significantly enhanced the mRNA levels of amylase, chymotrypsinogen, elastase 1, elastase 2 and carboxypeptidase. Under the treatment of activin A, sodium butyrate and dexamethasone, differentiated cells were polygonal in shape with large, round, and center-situated nuclei. According to the observation of immunofluorescence staining, amylase was positive expressed at the final stage. CONCLUSION: These data indicate that exocrine pancreatic differentiation of ES cells is induced by sodium butyrate, activin A and dexamethasone. The combination of pancreatic inducing factors improves the differentiating efficiency.     

Comparing fire spread algorithms using equivalence testing and neutral landscape models

We demonstrate a method to evaluate the degree to which a meta-model approximates spatial disturbance processes represented by a more detailed model across a range of landscape conditions, using neutral landscapes and equivalence testing. We illustrate this approach by comparing burn patterns produced by a relatively simple fire spread algorithm with those generated by a more detailed fire behavior model from which the simpler algorithm was derived. Equivalence testing allows objective comparisons of the output of simple and complex models, to determine if the results are significantly similar. Neutral landscape models represent a range of landscape conditions that the model may encounter, allowing evaluation of the sensitivity and behavior of the model to different landscape compositions and configurations. We first tested the model for universal applicability, then narrowed the testing to assess the practical domain of applicability. As a whole, the calibrated simple model passed the test for significant equivalence using the 25% threshold. When applied to a range of landscape conditions different from the calibration scenarios, the model failed the tests for equivalence. Although our particular model failed the tests, the neutral landscape models were helpful in determining an appropriate domain of applicability and in assessing the model sensitivity to landscape changes. Equivalence testing provides an effective method for model comparison, and coupled with neutral landscapes, our approach provides an objective way to assess the domain of applicability of a spatial model.     

Effect of simvastatin on expression of integrin-linked kinase and renal tubulointerstitium injury in rats induced by high fat diet

AIM: To explore the effect of simvastatin on expression of integrin-linked kinase (ILK) and the injury of renal tubulointerstitium in the rats induced by high fat diet. METHODS: Fifty-four 6-8 week-old female Wistar rats were randomly assigned to the following three groups: high fat diet group, simvastatin group (rats were fed with high fat diet plus 10 mg·kg-1·d-1 simvastatin) and control group. Six rats in each group were sacrificed at 4th week, 10th week, and the others at 20th week. The injury of renal tubulointerstitium was observed under microscope with HE staining and the expression of renal ILK was determined by Western blotting analysis and immunohistochemistry. Levels of total cholesterol (TC) and triglyceride (TG) in serum were measured by enzymatic colormetric methods. RESULTS: The serum TC and TG levels and the expression of renal ILK significantly increased in both high-fat diet group and simvastatin group, compared to control group at 4th week, reaching a maximum at 20th week (P<0.01). Tubulointerstitium injuries including vacuolar degeneration, syncytial change, clody swelling, necrosis and atrophy of renal tubular epithelial cells, thinning of tubal wall, lumens compensational expansion or even abolition, and inflammatory cell infiltration, interstitial fibrosis were found in both high fat diet group and simvastatin group, compared to control group at 4th week, worsened to a maximum at 20th week. However, all of these ameliorated in simvastatin group, compared to high fat diet group at each time point. CONCLUSION: The results indicate that high-fat diet induces significant lesion of renal tubulointerstitium and increased expression of renal ILK. Simvastatin may play an important role in protecting against tubulointerstitium injury induced by hyperlipoidemia by down-regulating the expression of renal ILK.     

大白菜叶片长宽性状的QTL定位

利用一套大白菜双单倍体（DH）群体和包含287个分子标记的遗传图谱,运用复合区间作图法对叶长、无柄叶长、有柄叶长、叶宽、叶柄长、叶柄宽6个叶部形态学性状进行QTL定位及遗传效应的分析。结果表明：在4个连锁群上共检测到12个QTL,分别为叶长相关的3个,无柄叶长相关的2个,有柄叶长相关的2个,叶宽相关的1个,叶柄长相关的2个,叶柄宽相关的2个。各位点的遗传贡献率介于7.5 %~18.9 %之间。其中,叶长与叶柄长的QTL存在共定位,叶宽与叶柄宽的QTL也存在共定位。     

钾素不同用量对莴苣硝酸盐积累的影响

以莴苣为试材,采用通用随机排列设计,利用基质培育方法,研究不同供应水平的钾素营养对莴苣硝酸盐积累的影响.结果表明:在供试条件下,硝酸盐含量与硝酸还原酶活性在一定浓度范围内都随着钾素浓度的增大而降低.     

几个鸢尾新品种栽培研究

对几种在郑州地区新引进的德国彩花鸢尾进行了栽培研究。结果表明:不朽白和黑骑士生长较旺,花朵较大,但开花率低,花期较短,繁殖系数低;黄娃娃和血石长势中庸,花朵中等,开花率高,花期较长,繁殖系数也高;金娃娃与黄链长势中庸偏下,花朵较小,但花期较长,金娃娃繁殖系数中等,黄链繁殖系数最低;紫边白植株矮小,长势较弱,花朵也小,但花期较长,繁殖系数较低。无论任何品种在郑州地区均能良好生长,且其生长、开花、繁殖均与土壤肥沃、疏松、排水良好呈正相关。