Quick detection of sex determining region protein gene in mouse fetuses

AIM: To detect quickly the Y-chromosome specific sex determining region protein (Sry) gene in mouse fetuses on embryonic day 14.5 with a PCR method. METHODS: We designed specific primers with the OLIGO 5. 0 software. Templates were prepared in 30 minutes by the following way. About 1 mg embryonic tissue but not fetal liver was suspended, and treated with 200μL of lysis buffer, consisting of PCR buffer containing 20 mg/L proteinase K, 0. 5% NP-40, and 0.05% Tween 40, at 60°C for 15 minutes, heated for 5 minutes at 100 °C, 10μL was used as template. The PCR react ion was performed in 50μL, using two sets of primers specific for Sry gene (chromosome Y) and IL-3 gene (chromosome 11) . PCR conditions and cycle numbers were optimized. The assessment of the results was done by electrophoresis in 3% agarose run at high voltage. The specificity of the method was conf irmed by fluorescent in situ hybridization (FISH) using a specific male probe on embryonic tissue cells. RESULTS: Electrophoresis showed that PCR product of male control DNA consisted of a 649 bp product representing the IL-3 gene and a 444 bp product representing the Y-specific Sry gene, female control DNA only one 649 bp product. Fetuses with two bands matching those as seen inmale control DNA are the presumpt ive male fetuses. Fetuses, only the IL-3-associated 649 bp band, are the presumptive female fetuses. These were confirmed by FISH. The ent ire procedure took <3. 5 h. CONCLUSION: The established PCR assay offers a quick, simple, accurate, and sensitive detection of sex determining region protein gene in mouse fetuses. This method allowed the preparation and culture of pure male and female hematopoietic stem cells from fetal tissue.     

梨新品种‘花盖王’

 ‘花盖王’是从‘花盖梨’中选出的梨四倍体芽变新品种。该品种除保持‘花盖梨’的优质、丰产、抗寒等优良性状外，最突出特点是果实大，单果质量是‘花盖梨’的2倍多，最大为250 g。     

Expression and role of CD134 and NF-κB in renal tissue of lupus nephritis

AIM:To investigate the role of CD134 (OX40) and NF-κB in the pathogenesis of lupus nephritis (LN).METHODS:Renal in situ CD134 and NF-κB expression were examined in 40 biopsy specimens from LN patients by immunohistochemistry and microwave-based immunohistochemistry, respectively. The relationship between expression of CD134 and NF-κB was analyzed.RESULTS:The expression of glomerular and tubular CD134 and NF-κB in LN were higher than that in normal control, especially in class Ⅳ LN, where there was intense staining of endothelial cell, distal tubules, and interstitial mononuclear cell. The CD134 expression of glomerular and tubular was closely related to NF-κB expression, respectively (r=0.5542, P＜0.05;r=0.6279, P＜0.05). CONCLUSIONS:The abnormal expression of costimulatory molecule CD134 was well evidenced in LN. Strong expression of renal in situ NF-κB was likely mediated by CD134 signal pathway, which may play an important role in the pathogenesis of LN.     

Effect of mutant β-catenin gene on the proliferation of hepatocytes

AIM:The β-catenin is a key molecule in the Wnt signal pathway, which plays a critical role in normal development and tumorigenesis. However, the mechanisms of the β-catenin on the cell growth control are still not completely defined. The aim of this study was to test the hypothesis that the mutant β-catenin may regulate the hepatocyte proliferation. METHODS: The immortalized murine hepatocyte cell line, AML12, was used for this study. A plasmid that contain mutant β-catenin S33Y was transfected into the AML12 cells and a stable cell line AML12S33Y was established. The cell growth property of this cell line and the parental cell were compared by flow cytometry analysis and direct cell count. The cells were also tested for the ability to form soft agar colonies, and the ability to form tumors in the severe immune deficient mice (SCID). RESULTS:1. The mutant β-catenin containing cell line AML12S33Y has higher proliferating index compared with the parental AML12 cells (P<0.01), suggesting that mutant β-catenin promotes cell growth. 2. The mutant β-catenin cells formed small colonies in soft agar after 4 weeks of culture, but did not generate tumor in SCID mice. CONCLUSION:The mutant β-catenin promotes liver cell growth.     

Effect of tea-polyphenols on the activation of NF-κB and the expression of TGF-β1 mRNA in THP-1 cells incubated with VLDL,LDL or ox-LDL

AIM:To investigate the effect of tea-polyphenols (TP) on the activation of NF-κB and the expression of TGF-β₁ mRNA in THP-1 cells (a human acute monocytic leukemia cell line). METHODS:THP-1 cells were incubated with the different concentrations of TP, VLDL, LDL or ox-LDL. In the THP-1 cellls, the nuclear malposition rate of NF-κB was detected with immunohistochemistry technique, the positive index of the TGF-β₁ mRNA expression was detected by hybridization in situ, and accumulation of total cholesterol (TC) in cells incubated with 0.4-40 μg/L TP was determined with oxidase assay. RESULTS:The nuclear malposition rate of NF-κB, the positive index of the TGF-β₁ mRNA expression and TC in THP-1 cells incubated with 0.4-40 μg/L of TP were lower than those with 0 μg/L of TP in TP-V group, TP-L group and TP-O (P＜0.05). The differences of these markers in THP-1 cells incubated with more than 40 μg/L TP in TP-V group, TP-L group and TP-O were not statistically significant, compared with TP-C group (P＞0.05). CONCLUSION:TP inhibited the activation of NF-κB, the expression of TGF-β₁ mRNA and the foam cell formation in the mono-macrophage.     

Predicting the spatial distribution of ground flora on large domains using a hierarchical Bayesian model

Accomodation of important sources of uncertainty in ecological models is essential to realistically predicting ecological processes. The purpose of this project is to develop a robust methodology for modeling natural processes on a landscape while accounting for the variability in a process by utilizing environmental and spatial random effects. A hierarchical Bayesian framework has allowed the simultaneous integration of these effects. This framework naturally assumes variables to be random and the posterior distribution of the model provides probabilistic information about the process. Two species in the genus Desmodium were used as examples to illustrate the utility of the model in Southeast Missouri, USA. In addition, two validation techniques were applied to evaluate the qualitative and quantitative characteristics of the predictions.This revised version was published online in May 2005 with corrections to the Cover Date.     

荔枝、龙眼园使用机油乳剂的安全性及其对荔枝蒂蛀虫的防治研究

利用机油乳剂对果树进行安全性和防治荔枝蒂蛀虫Conopomorpha sinensis Bradley研究试验，结果表明，喷施机油乳剂后荔枝和龙眼的落果率与对照树4无差异，喷清水的对照树其蒂蛀虫蛀果率比喷机油乳剂的处理树高，分别为150、200和250倍机油乳剂处理的2.8、3.0和2.7倍。在荔枝、龙眼园应用机油乳剂防治害虫不会导致落叶、落果、畸形叶和畸形果，也不会延缓果实的生长和成熟及降低单果重，而有利于控制介壳虫、螨类、蛀果类等害虫。机油乳剂可替代广谱性化学农药的使用，减少化学农药的污染，有利于果园天敌的生存、繁殖，提高生物多样性，改善果园生态环境。     

红肉脐橙果肉中主要色素的定性及色素含量的变化

 运用薄层层析法及二极管阵列检测器与高效液相色谱联用的方法, 确定红肉脐橙果肉中含有番茄红素和β- 胡萝卜素两种主要色素, 它们在液相色谱图中占总峰面积(计溶剂峰) 的73. 57 %～77. 78 % , 番茄红素占55. 36 %～58. 49 % , β- 胡萝卜素占18. 21 %～19. 93 %。利用反相高效液相色谱仪以及番茄红素和β- 胡萝卜素标样制作标准曲线, 测定果实发育及室温贮存过程中色素的变化动态, 结果表明, 该脐橙果肉中的番茄红素在成熟期含量最高, 达109. 67μg·g^-1DM, β- 胡萝卜素此时也达到最高值(15. 52μg·g^-1DM) ; 番茄红素的含量在采收后基本呈下降趋势(但采后30 d 有显著升高) , 说明果肉中存在类胡萝卜素采后合成的现象; 贮存4 个月后, 番茄红素含量急剧降低至最高时的1/ 10。     

苹果的3种优良砧木

介绍了“B9”、“CG80”、“75-7-1”三种苹果矮化钻木，并以生产上应用较多的M26作为对照，研究了其矮化程度、早果性、嫁接亲和力、树体产量等性状。     

红肉脐橙(Citrus sinensis L.)果肉中特征色素提取方法探索

红肉脐橙是目前为止惟一果肉呈粉红色或红色的脐橙,其呈色色素尚未见报道。为深入研究其果肉中特征色素的构成及在果实生长发育和贮存过程中色素含量的变化,本研究在对该色素初步定性的基础上对提取方法进行了探索。在分析红肉脐橙果肉中色素萃取液的紫外可见吸收光谱时发现,该色素有类胡萝卜素的特征吸收峰,将萃取液进行薄层层析则进一步证实其内含有番茄红素。在色素提取的前处理方式上,本研究认为冷冻干燥样品的浸提效果较真空干燥的样品稳定(变异系数小),且浸提率相差无几,所以选择冷冻干燥作为合适的样品前处理方式。色素浸提条件的正交实验结果表明,最佳浸提条件为50℃、1.5h内浸提4次,浸提的料液比为1:60。此外,浸提温度、浸提时间、浸提料液比和浸提次数对浸提率都有显著影响,影响最大的因素是浸提温度,最佳温度为50℃,随温度降低浸提率显著降低。其次是侵提时间的长短,最佳浸提时间为1.5h,延长或缩短0.5h都会使浸提率下降。浸提次数对浸提率的影响是最小的。