豫北山区“粮—草—羊”高效平衡养羊模式现状

从粮食作物、经济作物及饲草的种植,饲草料的加工调制与利用,全舍饲肉羊的养殖及杂交繁育,以及羊粪尿的资源化利用等方面介绍了豫北山区“粮—草—羊”高效平衡养羊模式发展现状,以期为促进豫北山区资源可持续利用、山区生态环境良性循环和社会经济可持续发展提供思路。     

Effect of beclin-1 gene silencing on TuAKe-injured gastric cancer SGC-7901 cells

AIM: To observe the effect of beclin-1 silencing by the technique of RNA interference on the injury of human gastric cancer SGC-7901 cell by Sheliugu extract (the extract from tuber of Amorphophallus konjac, TuAKe). METHODS: To knock down the expression of beclin-1 gene, SGC-7901 cells were transfected with lentiviral vector carrying beclin-1-shRNA. The beclin-1 gene knock-down and non-knock-down SGC-7901 cells were treated with TuAKe. The cell viability was analyzed by CKK-8 assay. The percentages of apoptotic cells were detected by flow cytometry. The expression of beclin-1 and LC3 was detected by Western blot. RESULTS: The beclin-1 gene silencing decreased the protein expression of beclin-1 and increased the protein expression of LC3 in the SGC-7901 cells, leading to the decrease in cell viability and the increase in apoptotic rate (P<0.05). TuAKe increased the protein expression of beclin-1 and LC3 in the SGC-7901 cells, and decreased the protein expression of LC3 in the SGC-7901 cells with beclin-1 gene silencing, thus inhibiting the cell viability and increasing the apoptotic rate (P<0.05). CONCLUSION: Beclin-1 gene silencing inhibits the activation of beclin-1-related signaling pathway in gastric cancer SGC-7901 cells, and aggravates the injury of cell viability induced by TuAKe.     

不同浓度1-MCP对柿果保鲜效果研究及灰色关联性分析

以恭城月柿为试材，采用精准相温（-0.5±0.3）℃结合不同浓度（0（CK）、1、3、5 ?滋L/L）1-甲基环丙烯（1-MCP）的处理方式，探索1-MCP最佳处理浓度及对柿果保鲜效果的影响。结果表明，不同浓度1-MCP处理均可保持柿果的感官品质、色泽及抗坏血酸（VC）含量，降低乙烯呼吸速率和丙二醛含量，从而达到延长贮藏期和维持柿果品质的目的，且随着贮藏时间的延长，保鲜效果更加显著，但是对可滴定酸（TA）含量及呼吸强度并无显著影响。灰色关联分析可知，4个处理的关联度分别为0.566、0.885、0.654、0.722，按大小排序为1 ?滋L/L 1-MCP＞5 ?滋L/L 1-MCP＞3 ?滋L/L 1-MCP＞CK，说明采用浓度为1 ?滋L/L 1-MCP处理柿果能够具有最佳贮藏效果，5 ?滋L/L 1-MCP次之，3 ?滋L/L 1-MCP效果较差。     

PPARγ mediates effects of diosgenin on proliferation and apoptosis in human glioblastoma U87MG cells

AIM:To investigate the effect of diosgenin (Dio) on the proliferation, apoptosis and expression of peroxisome proliferator-activated receptor γ (PPARγ) in human glioblastoma U87MG cells and its possible mechanism. METHODS:Human astrocytes (HA) and U87MG cells were cultured in vitro and treated with Dio (0, 10, 20, 30, 40 and 50 μmol/L) and GW9662 (5 μmol/L) for 48 h, and then the cell viability was detected by CCK-8 assay. Cell colony formation assay was used to assess the proliferation potential. Flow cytometry was used to analyze the cell cycle distribution and apoptosis. The mRNA expression level of PPARγ was measured by RT-PCR. Western blot was used to determine the protein levels of PPARγ, cyclin D1, cyclin E1, Bcl-2 and Bax. RESULTS:Dio had no significant influence on the viabi-lity of HA (P>0.05). However, Dio remarkably reduced the viability of U87MG cells in a dose-dependent manner (P<0.05) with IC₅₀ of 24.31 μmol/L. Meanwhile, Dio remarkably diminished colony formation ability (P<0.05), induced G₀/G₁ phase arrest of the cell cycle and apoptosis (P<0.05), up-regulated the expression of PPARγ at mRNA and protein levels, increased the protein level of Bax (P<0.05), and down-regulated the protein levels of cyclin D1, cyclin E1 and Bcl-2 (P<0.05) in a dose-dependent manner. However, these effects induced by Dio were inhibited by GW9662 (P<0.05), a specific inhibitor of PPARγ. CONCLUSION:Dio may inhibit proliferation and induce apoptosis in human glioblastoma U87MG cells most likely via up-regulating the expression of PPARγ, and then down-regulating the protein levels of cyclin D1, cyclin E1 and Bcl-2, and up-regulating the protein level of Bax.     

伐地那非增加EGCG-β-乳球蛋白纳米粒对肝癌细胞的抑制作用

较高浓度的EGCG才能抑制癌细胞的增殖,通过纳米化和EGCG与其他药物的联合使用是提高EGCG生物活性的重要策略。本研究将EGCG和伐地那非（VD）同时包埋于β-乳球蛋白（β-Lg）纳米载体中,制备出EGCG-VD-β-Lg纳米粒（EVβ-NPs）,体外试验证实,EVβ-NPs能提高人肝癌细胞（HepG2细胞）中Caspase-3活性,使HepG2细胞在S期产生明显的阻滞,诱发细胞核分裂,从而导致HepG2细胞凋亡。研究结果表明,将EGCG与微量的VD联合使用,并通过纳米化包埋可以显著提高EGCG的抗癌活性。这一方法在EGCG抗癌制品的开发方面具有潜在的价值。     

白叶枯病菌效应子XopN在拥有OsSWEET11同源基因的水稻品种上发挥毒性作用

【目的】黄单胞菌细胞外泌蛋白质(Xanthomonas outer proteins, Xops)是植物病原黄单胞菌高度保守的毒性效应子或毒性辅助组分,通过细菌Ⅲ型分泌系统分泌到细菌细胞外部,然后转入植物细胞而发挥病理作用。水稻黄单胞菌水稻致病变种即水稻白叶枯病菌的标准菌株PXO99^A分泌的XopN是一种毒性效应子,通过影响寄主免疫反应而使水稻发病。但是,XopN对病菌毒性的影响是否因水稻品种的不同而异,还有待研究。【方法】利用同源双交换技术,先敲除了PXO99^A的XopN基因,获得了∆XopN突变体,又经过遗传互补,得到了回补菌株∆XopN/XopN。通过营养肉汤液体培养,测定了XopN对病菌繁殖能力的影响;根据文献选用14个水稻品种,通过接种实验,测定了PXO99^A对这些品种的毒性与XopN敲除或回补的影响;在XopN发挥毒性作用的水稻品种上,测定了隐性抗病基因OsSWEET11/xa13与显性感病基因OsSWEET11/Xa13受病菌侵染而表达的情况,分析了XopN敲除或回补的影响。【结果】在营养肉汤液体培养过程中,病菌突变体菌株∆XopN的繁殖速度明显低于野生型PXO99^A。PXO99^A、∆XopN和∆XopN/XopN接种水稻后,根据其在水稻叶片组织内的繁殖量及随后产生的白叶枯病症状的严重程度,将供试的14个水稻品种分为两种情况。一是感病程度与病菌XopN是否敲除或回补无关,这有10个水稻品种（IRBB1、IRBB3、IRBB8、IRBB10、IRBB14、IR24、IRBB203、IRBB204、IRBB205和IRBB211）,PXO99^A、∆XopN和∆XopN/XopN对它们的毒性无明显差别。二是XopN对病菌毒性发挥作用的水稻品种,包括高度感病的3个品种（IRBB208、Asominori和日本晴）和低感品种IRBB13。与PXO99^A或∆XopN/XopN相比,∆XopN对这4个水稻品种的毒性大为降低。在IRBB13上,病菌侵染对隐性抗病基因OsSWEET11/xa13在叶片内的表达发生抑制作用,这一效应与XopN的毒性功能相关。相反,日本晴显性感病基因OsSWEET11/Xa13却受病菌侵染的诱导,在叶片内的表达水平大幅度提高。IRBB208和Asominori携带OsSWEET11/Xa13同源基因,该同源基因在叶片内的表达水平也因病菌侵染而大幅提高。在这4个水稻品种上,PXO99^A和∆XopN/XopN能够诱导OsSWEET11/Xa13或其同源基因表达,但∆XopN无此作用。另外,XopN对病菌在非寄主植物烟草上诱发过敏反应有量变贡献,相比PXO99^A或∆XopN/XopN,∆XopN引起过敏反应的程度有所降低。【结论】XopN是一个有限广谱性效应子,在拥有OsSWEET11同源基因的水稻品种上发挥毒性作用。XopN也是病菌繁殖所需要的,对病菌在非寄主植物上诱导过敏反应有一定贡献。     

玉米热激蛋白ZmHSP70-12基因的克隆及表达分析

热激蛋白70(HSP70)是原核和真核细胞中普遍存在的一种高度保守的分子伴侣。从玉米中克隆1个HSP70家族成员。该基因cDNA序列全长为1 992 bp,开放阅读框为2 352 bp,编码663个氨基酸,蛋白质分子量约75.0 kD。蛋白结构预测及同源比对分析表明,该基因编码蛋白含ATPase位点和HSP70保守结构域,与拟南芥AtHSP70-12序列高度相似,命名为ZmHSP70-12。蛋白亚细胞定位显示,ZmHSP70-12蛋白在内质网中表达。实时荧光定量PCR分析表明,ZmHSP70-12对非生物胁迫高温、干旱均具有明显的应答反应,推测ZmHSP70-12是玉米中与胁迫逆境相关的基因。     

Intensive organic vegetable production increases soil organic carbon but with a lower carbon conversion efficiency than integrated management

Intensive vegetable production in greenhouses has rapidly expanded in China since the 1990s and increased to 1.3 million ha of farmland by 2016, which is the highest in the world. We conducted an 11‐year greenhouse vegetable production experiment from 2002 to 2013 to observe soil organic carbon (SOC) dynamics under three management systems, i.e., conventional (CON), integrated (ING), and intensive organic (ORG) farming. Soil samples (0–20 and 20–40 cm depth) were collected in 2002 and 2013 and separated into four particle‐size fractions, i.e., coarse sand (> 250 µm), fine sand (250–53 µm), silt (53–2 µm), and clay (< 2 µm). The SOC contents and δ¹³C values of the whole soil and the four particle‐size fractions were analyzed. After 11 years of vegetable farming, ORG and ING significantly increased SOC stocks (0–20 cm) by 4008 ± 36.6 and 2880 ± 365 kg C ha^?1 y^?1, respectively, 8.1‐ and 5.8‐times that of CON (494 ± 42.6 kg C ha^?1 y^?1). The SOC stock increase in ORG at 20–40 cm depth was 245 ± 66.4 kg C ha^?1 y^?1, significantly higher than in ING (66 ± 13.4 kg C ha^?1 y^?1) and CON (109 ± 44.8 kg C ha^?1 y^?1). Analyses of ¹³C revealed a significant increase in newly produced SOC in both soil layers in ORG. However, the carbon conversion efficiency (CE: increased organic carbon in soil divided by organic carbon input) was lower in ORG (14.4%–21.7%) than in ING (18.2%–27.4%). Among the four particle‐sizes in the 0–20 cm layer, the silt fraction exhibited the largest proportion of increase in SOC content (57.8% and 55.4% of the SOC increase in ORG and ING, respectively). A similar trend was detected in the 20–40 cm soil layer. Over all, intensive organic (ORG) vegetable production increases soil organic carbon but with a lower carbon conversion efficiency than integrated (ING) management.     

miR-204 inhibits proliferation of Hodgkin lymphoma cells by targeting Sirt1/p53 signaling pathway

AIM: To investigate the effect of microRNA-204 (miR-204) on the proliferation of Hodgkin lymphoma cells and the underlying mechanism. METHODS: The expression of miR-204 and Sirt1 mRNA in Hodgkin lymphoma tissues was detected by RT-qPCR. After transfection with miR-204 mimic, Sirt1 siRNA and miR-204 mimic+pcDNA3.1-Sirt1 into the L428 cells, the cell viability and BrdU incorporation were measured by CCK-8 assay and BrdU assay, respectively. The protein levels of Sirt1 and acetylated p53 (ac-p53) were determined by Western blot.The targeting relationship between miR-204 and Sirt1 was verified by double luciferase reporter assay. RESULTS: The low expression of miR-204 and the high mRNA expression of Sirt1 were found in the Hodgkin lymphoma tissues. Compared with control group, the cell viability, BrdU incorporation and the protein levels of Sirt1 and ac-p53 were significantly decreased after L428 cells were transfected with miR-204 mimic or Sirt1 siRNA (P<0.05). Compared with miR-204 mimic alone group, the cell viability, BrdU incorporation and the protein levels of Sirt1 and ac-p53 were increased after L428 cells were co-transfected with miR-204 mimic and pcDNA3.1-Sirt1 (P<0.05). The results of double luciferase reporter assay confiermed that Sirt1 was the target gene of miR-204. CONCLUSION: The inhibitory effect of miR-204 on the proliferation of L428 cells may be achieved by inhibiting the expression of Sirt1 and promoting the up-regulation of ac-p53.