Effect of Yigu capsule drug-containing serum on apoptosis and tartrate resistant acid phosphatase secretion in rat osteoclasts

AIM:The effects of Yigu capsule on tartrate resistant acid phosphatase (TRAP) secretion and apoptosis in rat osteoclasts were investigated in order to further explore its mechanism of preventing and treating osteoporosis.METHODS:(1) Twenty-month-old Sprague-daweley rats were randomly divided into two groups(Yigu capsule group and saline group), and the drug-containing serum and control serum were prepared. (2) The newborn Sprague-daweley rat osteoclasts were cultured with different concentrations of Yigu capsule drug-containing serum. At different time point, TRAP activity was measured and the survival osteoclast was counted under reverse microscope.The percentage of osteoclast apoptosis was observed under fluorescence microscope after acridine orange staining.RESULTS:TRAP activity was lower and the percentage of osteoclast apoptosis was higher in drug-containing serum group than in control group at 24, 48 and 72 h(P<0.01), respectively, and the survival osteoclasts were less in drug-containing serum group than in control groups at 24, 48 and 72 h(P<0.01).CONCLUSIONS:These data suggest that Yigu capsule drug-containing serum induces apoptosis and inhibits TRAP activity in osteoclasts, which may be one of the mechanisms of Yigu capsule preventing and treating osteoporosis.     

Expression of cyclooxygenase-2 in IL-1β-stimulated mesangial cells is medicated by NF-κB/IκB signal pathway

AIM: To investigate the role of NF-κB/IκB signal pathway in the regulation of cyclooxygenase-2 (COX-2) expression in human mesangial cells (HMC). METHODS: The PGE₂ concentration in supernatants of HMC was measured by radioimmunoassay. COX-2 mRNA and protein expression were determined by RT-PCR and Western blot. Electrophoretic mobility shift assay (EMSA) and Western blot were used to detect the activity of NF-κB and degradation of IκB. RESULTS: IL-1β significantly upregulated COX-2 expression and PGE₂ production in HMC. Significant up-regulation of NF-κB activation, nuclear translocation of p65 subunit, and degradation of IκB α and IκB β were observed in IL-1β-induced HMC. CONCLUSION: Expression of COX-2 in IL-1β-induced HMC is mediated by NF-κB/IκB signal pathway.     

Effect of YIGU capsule on IGF-I mRNA and protein expression in rat osteoblasts in vitro

AIM: The effects of YIGU capsule on proliferation and IGF-I mRNA protein expressions in osteoblasts were studied. METHODS: (1) Forty 12-month old Sprague-Dawley female rats were divided randomly into four groups (YIGU capsule high dose group, medium dose group and low dose group; saline group), the drug-containing serum and control serum were prepared. (2) The new-born Sprague-Dawley rat osteoblasts were cultured with different YIGU capsule drug-containing serum at different concentrations and different exposure time. MTT method was used to observe proliferation of osteoblasts. (3) RT-PCR method was used to measure the relative IGF-I mRNA levels and ELISA method was used to measure IGF-I secretion at different exposure time. (4) ELISA method was used to measure IGF-I secretion at different exposure time. RESULTS: (1) Proliferation of osteoblasts was more than the control groups after 48, 72 and 96 h, respectively (P<0.01); (2) The relative IGF-I mRNA levels and IGF-I protein expression were higher than those in control group after 48, 72 and 96 h, respectively (P<0.01 or P<0.05). CONCLUSIONS: It was suggested that YIGU capsule drug-containing serum promoted proliferation, IGF-I mRNA and protein expression. These results may be parts of the mechanisms of YIGU capsule to prevent and treat osteoporosis.     

‘紫罗兰’白菜叶片再生体系的建立

以‘紫罗兰’白菜无菌苗叶片为外植体,研究了不同6-BA和NAA浓度组合对白菜真叶不定芽再生的影响,同时采用不同生根培养基对再生芽进行生根诱导。结果表明:诱导‘紫罗兰’白菜真叶分化的适宜培养基为MS+2mg/L 6-BA+1mg/L NAA+4mg/L AgNO3,每个外植体平均再生芽数为1.42个;生根培养基以MS+0.5mg/L NAA最佳,平均每个不定芽诱导根数为16.54个。该试验结果为‘紫罗兰’白菜材料的快速繁殖提供了理论和技术参考。     

白藜芦醇和褪黑素对绵羊胎儿成纤维细胞转染及卵母细胞体外成熟的影响

试验旨在利用白藜芦醇(Re)和褪黑素(MT)提高绵羊胎儿成纤维细胞电穿孔转染(电转)效率、优化卵母细胞体外成熟体系,以提高转基因动物生产效率。将绵羊胎儿成纤维细胞分为6组:对照组、添加白藜芦醇高、中、低(R^-5、R^-6、R^-7)3种浓度组及白藜芦醇和褪黑素联合添加组(R^-5M^-5、R^-6M^-7),通过电转效率和细胞凋亡情况确定电转体系优化方案;通过对细胞周期、线粒体膜电位及活性氧(ROS)的检测研究其作用机制。将绵羊卵母细胞分为对照组及白藜芦醇和褪黑素(R^-5M^-5、R^-6M^-7)联合添加组,通过绵羊卵母细胞体外成熟后卵丘细胞扩展、第一极体排出及体外受精胚胎的卵裂率检测其对卵母细胞体外成熟的作用。研究结果显示,分离的绵羊胎儿成纤维细胞P3代增殖旺盛,细胞生长曲线呈典型的S型;与对照组相比,所有试验组胎儿成纤维细胞的电转效率均显著提高(P<0.05),R^-5M^-5组电转效率最高;R^-6M^-7组显著降低细胞凋亡率(P<0.05),R^-5、R^-6、R^-5M^-5和R^-6M^-7组均显著促进细胞增殖(P<0.05);R^-6、R^-7、R^-5M^-5和R^-6M^-7组G1期细胞增加,S期细胞相对减少,但差异均不显著(P>0.05);R^-6、R^-7、R^-5M^-5和R^-6M^-7组成纤维细胞内ROS水平均显著降低(P<0.05),R^-6、R^-5M^-5和R^-6M^-7组线粒体膜电位均显著增加(P<0.05);R^-6M^-7组显著促进绵羊卵丘细胞扩展、提高卵母细胞第一极体排出率和体外受精胚胎发育能力(P<0.05)。综上,白藜芦醇和褪黑素可以优化绵羊胎儿成纤维细胞电转体系,提高卵母细胞体外成熟效率,为白藜芦醇和褪黑素在转基因动物生产中的应用提供参考依据。     

类器官的研究进展及应用前景

类器官（organoids）来源于自组织和自我更新的干细胞，是利用干细胞的自组织特性进行体外3D培养后形成的细胞团，与来源器官密切相关，再现了来源器官的三维细胞结构，并为探索来源器官的发病机制提供了新的模型。类器官系统是由自分泌、旁分泌或邻分泌信号调节下的细胞，或者外源性添加的细胞外基质（extracellular matrix，ECM）底物、小分子和生长因子等衍生而来，这些因素的相互作用创造了一个动态的环境，指导干细胞的自我更新和分化，以及细胞在类器官中的自我组装。诱导多能干细胞（induced pluripotent stem cells，iPSC）重编程方法结合3D类器官工具，使患者来源的类器官作为动物模型和人类临床试验之间的桥梁，是对细胞研究和在体试验的补充。在研究来源器官发育、生物学和病理生理学方面，类器官不仅是一种比传统细胞培养更具生理相关性的体外模型，而且还是再生医学和个性化医学领域中的新模型，有望成为研究营养素、药物、毒物及毒素等的作用机制及药物的筛选、再生医学等领域的重要模型。总之，类器官技术的发展增强了人们对器官和组织生理生化功能的认知。作者对肠、脑、肺脏、肝脏、子宫、卵巢等类器官培养和应用的研究进展进行综述，以期为类器官相关科研及应用提供参考。     

低温浸渍对“玫瑰香”低醇桃红葡萄酒品质的影响

以天津汉沽产区的"玫瑰香"葡萄为试材,分别选取了2、6、10℃为酒精发酵前浸渍温度,并以24、48、72h为低温浸渍时间,研究了低温浸渍前处理方式对低醇桃红葡萄酒质量的影响,并通过固相微萃取结合气质联用法对其香气物质进行了定量分析。结果表明:6℃下低温浸渍处理葡萄汁48h后再发酵获得的低醇葡萄酒理化指标和感官指标均优于其它浸渍温度和时间。在此条件下酿的低醇葡萄酒香气物质含量明显高于对照组,其颜色浅桃红、果香浓郁、清新爽口、口感均衡、余味较长,具有"玫瑰香"葡萄的典型性。其中对22种香气成分进行了定量分析,发现低温浸渍增加了总的香气成分含量,赋予了低醇葡萄酒更多的新鲜水果香和花香,一定程度上弥补了由于低醇发酵造成的香气不足。说明短期低温浸渍可以改善"玫瑰香"低醇桃红葡萄酒的品质。     

红叶紫薇“一步法”快速繁殖试验

以红叶紫薇当年生新枝具腋芽茎段为外植体材料,研究其灭菌时间长短对外植体污染率及腋芽萌发率的影响,根据基本培养基上其离体腋芽萌发诱导情况,对培养基(MS,1/2MS,WPM,1/2WPM)进行筛选,并根据植物生长调节剂(BA,NAA,KT)及其质量浓度的不同配合,筛选出无菌苗"一步法"增殖、生根培养基。试验结果显示:使用0.1%Hg Cl2对外植体灭菌12 min,能在保证低污染率(13.33%)的同时,获得较高的腋芽萌发率(80.00%);1/2MS培养基上的腋芽萌发率(96.67%)和腋芽长度(1.60 cm)均显著高于其他3种培养基;在1/2MS+1.0 mg/L BA+0.1mg/L KT培养基中,无菌芽可同时增殖及生根,芽增殖系数(4.67)、苗高(2.53 cm)、生根率(100%)及根数(4.33条)均显著高于其他处理。     

不同干燥方法对牡丹草化学成分及抗氧化活性的影响

以牡丹草为试材,采用晒干、阴干、硅胶干燥等方法进行干燥处理,通过紫外可见分光光度法测定各样品中总皂苷、总黄酮及多糖含量,研究不同干燥处理方法对牡丹草化学成分的影响,并以清除DPPH自由基(DPPH·)、ABTS自由基(ABTS+·)、羟基自由基(·OH)、超氧阴离子(O2·-)等多种抗氧化能力为指标,评价不同干燥方法处理后各牡丹草样品的抗氧化活性,同时结合Pearson相关性分析,探讨各样品抗氧化能力与样品中总皂苷、总黄酮及多糖含量的关系,进一步研究不同干燥处理方法对牡丹草抗氧化活性的影响,以期为牡丹草药材的炮制加工提供参考依据.结果 表明.70℃干燥处理后的牡丹草总黄酮、总皂苷和多糖含量最高,分别为0.0098、0.0088、0.0147 g·g-1,所测定的5种抗氧化指标中,70℃干燥处理后的牡丹草药材对O2·-、ABTS+·清除能力最强,IC50分别为4.981、3.543g·L-1;真空冷冻干燥处理后的牡丹草药材对DPPH·、·OH清除能力最强,其IC50分别为3.358、4.336 g·L-1;70℃干燥处理后的牡丹草药材总还原能力最强.通过Pearson相关性分析结果可知,牡丹草药材中的总皂苷及总黄酮含量与抗氧化能力存在显著相关性(P＜0.05).     

水杨酸浸种对谷子萌发期抗旱性的影响

为研究水杨酸对谷子萌发期抗干旱胁迫能力的影响,以衡谷15、衡谷13、冀谷19、衡饲1号谷子种子为材料,以18％的PEG模拟干旱环境,分别用7种浓度的水杨酸浸种24 h,测定发芽率、发芽指数、活力指数、根长、苗高、鲜重等,综合分析4种谷子的抗干旱胁迫能力.结果 表明:在干旱胁迫下,水杨酸浸种后各品种表现不同,有些品种的抗旱能力没有明显变化.而冀谷19和衡谷15的发芽能力显著提高,衡谷15、冀谷19、衡谷13的活力指数显著增加,0.6 mmol/L水杨酸浸种是最适浓度.通过隶属函数的综合分析可知水杨酸浸种后4个品种谷子抗旱性的大小为:衡谷13＞冀谷19＞衡饲1号＞衡谷15.