不同炮制方法对菟丝子总黄酮和多糖含量及抗氧化能力的影响

为研究不同炮制方法对菟丝子总黄酮和多糖含量、抗氧化能力的影响,采用水和醇对不同炮制方法制得的菟丝子进行提取,然后分别测定水和醇提取液中总黄酮和多糖含量、抗氧化能力。结果显示,与水洗晒干法相比较,雷氏法、酒炙法、粉碎水煮法等炮制方法显著地提高了菟丝子总黄酮和多糖的溶出能力,从而增加了菟丝子的抗氧化能力。醇提液中的总黄酮和多糖含量、抗氧化能力较水提液增加明显。另外,经过破碎处理的菟丝子中的总黄酮和多糖含量、抗氧化能力明显增多增强。结果表明,雷氏法、酒炙法、粉碎水煮法结合破碎处理和醇提更适合于菟丝子活性物质提取和利用。     

miR-129-3p targets Smad3 to inhibit viability and migration of NIH3T3 cells during TGF-β-induced transformation into myofibroblasts

AIM: To explore the effects of microRNA-129-3p (miR-129-3p) on the viability and migration of NIH3T3 cells during transforming growth factor-β (TGF-β)-induced transformation into myofibroblasts and the underlying molecular mechanisms. METHODS: RT-qPCR was used to examine the relative expression of miR-129-3p in renal cell carcinoma (RCC)-adjacent tissues and fibrotic renal tissue. NIH3T3 cells were stimulated with TGF-β to transform into myofibroblasts, and miR-129-3p expression level was detected. After transfection with miR-129-3p mimics for 48 h in vitro, the cell viability was measured by MTT assay, the protein expression level of Ki-67 was determined by Western blot, and the cell migration was observed by wound healing assay. The direct target of miR-129-3p was predicted by online database TargetScan and confirmed by dual-luciferase reporter assay. The expression level of target protein was further confirmed by Western blot. RESULTS: Compared with the RCC-adjacent tissues, the expression of miR-129-3p was down-regulated in fibrotic renal tissue (P<0.01). In TGF-β-induced NIH3T3 cell transformation into myofibroblasts, the expression of miR-129-3p was also decreased (P<0.01). Transfection with miR-129-3p mimics followed by TGF-β stimulation in the NIH3T3 cells inhibited the viability, Ki-67 expression and migration. TargetScan analysis showed miR-129-3p had binding sites in the 3'-UTR of Smad3, which was confirmed by dual-luciferase reporter assay. The results of Western blot further confirmed that miR-129-3p affected the expression of Smad3. CONCLUSION: miR-129-3p inhibits the viability and migration ability of NIH3T3 cells during TGF-β-induced transformation into myofibroblasts by directly targeting Smad3.     

miR-105 inhibits cell proliferation,migration and invasion abilities in non-small-cell lung cancer cells by targeting and negative regulation of KIFC1

AIM:To study the effects of microRNA-105(miR-105) on the cell proliferation, migration and invasion abilities of non-small-cell lung cancer (NSCLC) H460 cells, and further to explore its mechanism. METHODS:The expression of miR-105 and kinesin family member C1 (KIFC1) mRNA in the NSCLC tissues and adjacent tissues and cells was detected by RT-qPCR. The protein expression of KIFC1 in the NSCLC tissues, adjacent normal tissues and cells was determined by Western blot. The H460 cells were divided into miR-105 group (transfection with miR-105 mimics), miR-negative control (NC) group (transfection with miR-NC), inhibitor-NC group (transfection with NC of inhibitor), inhibitor-miR-105 group (transfection with miR-105 inhibitor), si-NC group (transfection with NC siRNA), si-KIFC1 group (transfection with KIFC1 siRNA), miR-105+vector group (miR-105 mimics and pcDNA 3.1 co-transfection) and miR-105+KIFC1 group (miR-105 mimics and pcDNA 3.1-KIFC1 co-transfection). The cell proliferation was measured by MTT assay and colony formation assay. The migration and invasion abilities were detected by Transwell methods. The relative luciferase acitivity was evaluated by double luciferase reporter assay. RESULTS:Compared with the adjacent tissues, the expression of miR-105 was significantly decreased and the expression of KIFC1 was significantly increased in NSCLC tissues (P<0.05). Compared with human normal embryonic lung fibroblasts MRC-5, the expression of miR-105 in the H460 cells was significantly decreased, and the expression of KIFC1 was significantly increased (P<0.05). miR-105 inhibited the relative luciferase activity of H460 cells with wild-type KIFC1 and negatively regulated the protein expression of KIFC1. Over-expression of miR-105 and knockdown of KIFC1 expression significantly inhibited the proliferation, migration and invasion abilities of H460 cells. Over-expression of KIFC1 reversed the inhibitory effect of miR-105 on the cell proliferation, migration and invasion abilities of H460 cells. CONCLUSION:miR-105 inhibits the proliferation, migration and invasion abilities of NSCLC cells. The mechanism may be related to targeting and negatively regulating expression of KIFC1.     

雄性不育野生茶树开花生物学特性

为观察野生茶树雄性不育株开花生物学特性,探讨野生茶树雄性不育表型特征。以野生茶树雄性不育株及可育株为试材,田间观察了野生茶树的开花物候期、单花发育进程和花器形态特征,并采用离体萌发法、染色法测定其花粉活力和柱头可授性。结果表明：野生茶树雄性不育株开花物候期为9月上旬—10月上旬,全花期持续约29~33天,花期与对照材料相近。雄性不育花从花芽分化至花瓣凋谢平均历时约107天,比对照长14天。雄性不育花器结构具有典型的雄不育特征,表现为花冠开展度小,花丝弯曲畸形,花药彼此粘连,皱缩干瘪,不裂药,花药内无花粉或有微量败育花粉。雄性不育花雌性器官发育正常,柱头从开花前1天至开花后4天具有可授性,最佳授粉时间为开花后1天。所调查的野生茶树雄性不育类型属于花药败育型和花粉败育型。     

Is the Organic System Economically Viable? The Case of Pineapple in India’s Northeast

This study intends to fill the gaps in the organic agriculture literature emerging from infrequent use of farm budget-related data, overlooking the imputed costs and non-existence of long-run return and profitability estimates. The results suggest that the organic system is economically unviable as the growers fail to recover their total production cost. However, growers’ returns exceed their ‘perceived’ production cost, which excludes their imputed expenses. Temporal, spatial, and size-based analysis also attests to this. On the contrary, cultivation in two sample villages where the household’s size, its head’s age, percentage of land, and yield are significantly higher, remain economically viable.     

不同贮藏年限敖汉苜蓿种子活力及生理特性的研究

种子在贮藏过程中随着贮藏时间的延长出现老化现象,使种子活力下降、生理特性发生变化,从而降低种用价值.为探讨不同贮藏年限紫花苜蓿种子的活力变化,选择室温贮藏0～3年的敖汉苜蓿种子,比较分析种子劣变规律及生理特性的差异.结果表明:收获当年的敖汉苜蓿种子具有较高的硬实率,其抗逆性强,SOD、POD、CAT、MDA和脯氨酸含量最高,而发芽势和可溶性蛋白含量最低.贮藏起初2年,种子硬实率、不正常种苗数、死种子数、SOD、POD、CAT、MDA和脯氨酸含量降低,发芽率、发芽势和可溶性蛋白含量增加.随着贮藏时间的延长,种子可溶性蛋白含量降低,SOD、POD、CAT、脯氨酸和MDA积累量增加,种子质量下降,不正常种苗数和死种子数增多,种子活力下降.综合试验结果,贮藏2年有利于打破种子休眠,其种子质量最佳,种子发芽能力最强,但是种子抗逆性不高;贮藏1年存在轻度休眠,应进行打破硬实处理;贮藏3年的种子活力下降,其种用价值降低,不利于在人工草地建植中使用,生产中应贮藏2年后再播种效果较好.研究结果能较好地说明苜蓿种子在贮藏期间的活力变化,为我国人工草地建设中合理使用苜蓿种子提供理论依据.     

Paeonol affects viability and migration ability of hepatocellular carcinoma cells by up-regulating miR-424-3p and inhibiting PI3K/AKT signaling pathway

AIM To investigate the effect of paeonol on the viability and migration ability of hepatocellular carcinoma cells and its molecular mechanism. METHODS Human hepatocellular carcinoma Hep3B cells was treated with paeonol at different concentrations (50, 100, 200 and 400 mg/L). The cell viability was measured by CCK-8 assay to determine the optimal drug concentration. The Hep3B cells were divided into normal control (NC) group, paeonol group, miR-NC group, miR-424-3p group, paeonol+anti-miR-NC and paeonol+anti-miR-424-3p group. The expression level of miR-424-3p was detected by RT-qPCR. The migration ability was detected by Transwell assay. The protein levels of cyclin D1, matrix metalloproteinase 2 (MMP2), MMP9 and PI3K/AKT signaling pathway-related molecules were determined by Western blot. RESULTS Paeonol intervention inhibited the viability of Hep3B cells in a concentration-dependent manner (P<0.05). The concentration of paeonol at 200 mg/L was selected for the following study. Paeonol intervention inhibited the protein expression of MMP2 and MMP9 in the Hep3B cells, and inhibited the migration ability of the Hep3B cells. Paeonol intervention promoted the expression of miR-424-3p in the Hep3B cells (P<0.05). Over-expression of miR-424-3p inhibited the expression of cyclin D1, MMP2 and MMP9 in the Hep3B cells and inhibited cell viability and migration ability (P<0.05). Inhibition of miR-424-3p reversed the effect of paeonol on the viability and migration ability of the Hep3B cells (P<0.05). Paeonol inhibited phosphorylation levels of PI3K and AKT in the Hep3B cells and inhibited the activation of PI3K/AKT signaling pathway (P<0.05). Inhibition of miR-424-3p reversed the effect of paeonol on PI3K/AKT signaling pathway in the Hep3B cells (P<0.05). CONCLUSION Paeonol inhibits the viability and migration ability of hepatocellular carcinoma cells by up-regulating miR-424-3p and inhibiting PI3K/AKT signaling pathway.     

High expression of BIRC5 enhances cell viability,inhibits apoptosis and is associated with poor prognosis in gastric cancer

AIM To investigate the expression of baculoviral inhibitor of apoptosis protein repeat-containing protein 5 （BIRC5） in gastric cancer tissue and its relationship with prognosis of gastric cancer patients, and to explore the effect of BIRC5 knock-down on the viability and apoptosis of gastric cancer cells. METHODS The expression of BIRC5 was detected by immunohistochemistry in 67 cases of gastric cancer tissues and paracancerous tissues for analyzing the relationships with clinicopathological characteristics. The mRNA and protein expression levels of BIRC5 in gastric carcinoma cell lines （AGS, MKN-1 and MGC-803） and normal gastric epithelial cell line GES-1 were detected by RT-qPCR and Western blot. The AGS cells were divided into blank group （no treatment）, Ctr-sh group （blank plasmid transfection） and BIRC5-sh group （BIRC5-shRNA plasmid transfection）. The interference efficiency of BIRC5-shRNA was evaluated by Western blot. The cell viability was measured by MTT assay, the apoptosis was analyzed by flow cytometry, and the levels of apoptosis-related proteins cleaved caspase-3, Bax and Bcl-2 were determined by Western blot. RESULTS BIRC5 was mainly expressed in cytoplasm, and the positive expression rate of BIRC5 in the gastric cancer tissues was higher than that in the adjacent tissues （P<0.01）. The positive rates of BIRC5 in the gastric cancer patients at TNM Ⅲ~Ⅳ stages and with lymph node metastasis were higher than those in the patients at TNM Ⅰ~Ⅱ stages and without lymph node metastasis, respectively （P<0.05）. The survival time of the patients with positive BIRC5 expression was shorter than that of the patients with negative BIRC5 expression （P=0.011 2）. The cell viability in BIRC5-sh group was lower than that in blank group and Ctr-sh group at time points of 48, 72 and 96 h. The apoptotic rate in BIRC5-sh group was increased compared with blank group and Ctr-sh group. The protein levels of cleaved caspase-3 and Bax in BIRC5-sh group were higher than those in blank group and Ctr-sh group, while the protein expression of Bcl-2 in BIRC5-sh group was lower than that in blank group and Ctr-sh group （P<0.05）. CONCLUSION High expression of BIRC5 in gastric cancer indicates poor prognosis. BIRC5 promotes the growth of gastric cancer cells and inhibits apoptosis.     

莴苣种子贮藏条件研究

运用适当的种子起始含水量、贮藏温度和包装方式,可经济有效地保持种子的发芽率。种子的生活力与超氧化物歧化酶、酸性磷酸酶的活性呈正相关。     

梨43个品种花粉生活力及4种测定方法的比较

用花粉离体培养法测定比较了43个梨品种花粉离体萌发率和花粉管生长长度;从中选择花粉相对生活力低、中、高3组6个品种(今村秋和雪花梨、金二十世纪和丰水、青松和满天红)的花粉,采用I-KI染色法、蓝墨水染色法、MTT染色法、过氧化物酶染色法4种花粉生活力测定方法,以花粉离体萌发法为对照。结果表明,花粉培养24h后,43个梨品种的花粉萌发率平均为66.15%,其中花粉萌发率≥60%的品种约占84%,满天红萌发率最高为82.79%;花粉管长度平均为1.77mm,花粉管长度≥1mm的品种约占89%,其中奥萨二十世纪最长为3.25mm,说明多数梨品种花粉生活力较强。与对照相比,4种花粉生活力测定方法中I-KI染色法的测定值极显著偏低,MTT(噻唑蓝)染色法的测定值显著偏高,二者不适于梨花粉生活力的测定;过氧化物酶染色法测定值与对照差异显著,也不适宜用来测定梨花粉生活力;蓝墨水染色法测定值与对照没有显著性差异,可用于梨花粉生活力的快速测定。