柴胡愈伤组织提取物抗肝癌效用研究

目的:对中药柴胡的愈伤组织进行培养,并对其抗肿瘤活性进行初步研究。方法:提取柴胡愈伤组织活性成分(BSW-ws),对培养的肝癌细胞SMMC-7221给药,给药浓度分别为:10,20,30,40,60,80,100μg·mL~(-1);再用MTT法检测BSW-ws的细胞毒活性,并经流式细胞仪检测BSW-ws对细胞凋亡的影响。结果:(1)MTT法测BSW-ws的细胞毒活性结果显示,当给药浓度为60μg·mL~(-1)时达到IC50值;(2)流式细胞仪检测结果表明,BSW-ws能够促进肝癌细胞SMMC-7221的凋亡。结论:柴胡愈伤组织提取物BSW-ws通过促进癌细胞凋亡的形式抑制肝癌细胞SMMC-7221的增殖。     

肉苁蓉愈伤组织的超低温保藏方法

提出了一个优化的简单的超低温保藏方法,成功地运用于肉苁蓉愈伤组织的低温保藏. 为了获得最佳的实验结果,肉苁蓉愈伤组织首先在添加了6%二甲亚砜的B5培养基中进行预培养,然后用玻璃化保护剂在25℃处理20 min,最后投入液氮中进行冷冻. 玻璃化保护剂的成分为30%(j)甘油+15%(j)乙二醇+10%(j)二甲亚砜+0.5 mol/L蔗糖. 冷冻后的愈伤组织在30℃的水浴中迅速解冻,接着用25℃的1.0 mol/L蔗糖溶液洗净愈伤组织上附着的玻璃化保护剂,最后在B5培养基上对愈伤组织进行恢复性培养. 经过上述冻存处理的肉苁蓉愈伤组织存活率可达86％. 恢复培养5个月后,愈伤组织中苯乙醇糖甙类化合物的含量和产量分别达到冷冻前的97%和95%.     

苦参愈伤组织诱导及其杀虫抑菌生物活性测定

通过以苦参越冬芽、茎、子叶和幼叶为外植体诱导愈伤组织,四者在合适的培养基上均能诱导出愈伤组织,幼叶和子叶易于诱导愈伤组织。苦参叶和子叶诱导的愈伤组织处理淡色库蚊的死亡率为100％,处理桃蚜的死亡率分别为81．01％和85．42％,高于其他愈伤组织的活性;各愈伤组织处理朱砂叶螨的死亡率均达到了95％以上,与种子提取物的活性没有显著差异。只有子叶诱导的愈伤组织对油菜菌核病菌表现出了抑制作用,各愈伤组织对小麦赤霉病菌和黄瓜灰霉病菌均未表现出明显的活性,显著低于苦参种子提取物的活性。     

女贞子胚培养及愈伤组织诱导

采用不同培养基对女贞子的胚进行培养,并对其幼苗的叶片和嫩茎进行愈伤组织诱导.结果表明,在胚的萌发诱导中,以MS+1.00 mg·L-1 6-BA培养时,女贞子幼苗的叶长和根长最长;以MS+1.50 mg·L-1 6-BA培养时,女贞子幼苗的茎高最高;MS+1.50 mg·L-1 6-BA+0.30 mg·L-1 2,4...     

蓖麻愈伤组织对铜的抗性研究

为了探讨蓖麻愈伤组织对铜的抗性．研究了铜胁迫下蓖麻愈伤组织的增长及其铜吸收作用。结果显示,铜浓度为60mg·L^-1时,愈伤组织的增长受到抑制,抗性指数仅为33．87％;铜浓度为40mg·L^-1时,愈伤组织呈淡黄色,生长较快,抗性指数达到61．29％,并且这种抗性可以保持至连续继代培养6周之后。培养至第4周,铜浓度（mg·L^-1）为10、20、30、40各处理的抗性指数都高于第3周．各处理愈伤组织铜含量（mg·g^-1）依次为0．33、0．54、1．16、1．40。培养基铜浓度为40mg·L^-1。可作为筛选铜抗性蓖麻愈伤组织的临界值。     

仙客来愈伤组织诱导和不定芽形成

以仙客来品种“国旗红”成株的叶片和叶柄为外植体,分别接种到MS培养基(Murash ige&skoog)+6苄基嘌呤(BA)2.0(mg/L,下同)+萘乙酸(NAA)0.5和1/2MS+BA 2.0+激动素(KT)0.25+2,4二氯苯氧乙酸(2,4 D)0.5两种培养基,研究外植体类型、培养基、切割方式和摆放方法对愈伤组织诱导和不定芽形成的影响。结果表明,叶片是诱导愈伤组织和形成不定芽的理想外植体;将带主脉的叶片,以反面向上的方法接种到1/2MS+BA 2.0+KT 0.25+2,4 D 0.5中诱导效果最佳,愈伤组织诱导率达100%,不定芽形成率和平均个数分别为66.67%和4.0。此外,对不定芽的继代培养进行研究,结果表明,不定芽可继代增殖。     

稀土元素对水母雪莲细胞生长及黄酮类化合物合成的影响

研究了稀土元素钕(Nd3+)、铈(Ce3+)、镧(La3+)和混合稀土(MRE)对摇瓶液体培养的水母雪莲细胞生长及黄酮类化合物合成的影响. 发现Ce3+和La3+及混合稀土可促进雪莲细胞的生长及黄酮类化合物的合成,其中以Ce3+效果最佳. 当初始浓度为0.025 mmol/L的Ce3+添加到改良的MS培养基中时,细胞生物量和黄酮类化合物产量最高,分别可达17.7 g/L及942 mg/L,分别是不添加稀土元素的对照实验的134.4%和166.7%;同时发现在培养基中不含外源激素6-BA的条件下,合适浓度的Ce3+可替代6-BA对雪莲细胞生长及黄酮类化合物合成的促进作用,而在培养基中不含NAA时,Ce3+不能替代NAA对雪莲细胞生长及黄酮类化合物合成的促进作用.     

NAA与BA对康乃馨愈伤组织诱导的影响

文章主要研究了NAA与BA对康乃馨愈伤组织诱导的影响,结果表明：单独使用NAA或BA时,能够诱导出愈伤组织;NAA浓度为0．2mg/L,BA浓度为2mg／L时,产生愈伤组织比例最高,生长状态最好。     

藏红花愈伤组织诱导和褐化抑制

对藏红花的球茎、无菌芽和幼叶的愈伤组织诱导条件进行研究,同时探讨愈伤组织诱导过程中的褐化和污染问题．结果表明：与球茎相比,以无菌芽和幼叶为外植体,愈伤组织诱导率较高,诱导时间较短,但形成的愈伤组织易褐化．较大球茎（≥20g）也是藏红花愈伤组织诱导较好的外植体,愈伤组织诱导速度随球茎储藏时间的增加而升高．含有2．0mg／L2,4-二氯苯氧乙酸（2,4-D）和O．5mg／L6-苄氨基嘌呤（6-BA）的MS（Murashige and Skoog）培养基有利于球茎愈伤组织的诱导,30d愈伤组织诱导率可达70％以上．在愈伤组织诱导过程中,连续转接外植体至新鲜培养基可以有效降低褐化率,而在培养基中添加活性炭（AC）和聚乙烯吡咯烷酮（PVP）不能有效抑制褐化．表面灭菌前采用流水冲洗过夜可有效减少球茎表面附生菌,明显降低污染率．     

藏雪莲水母雪兔子中矿质元素的测定与分析

经HNO3/H2 O2湿法微波消解制样,利用全谱直读电感耦合等离子原子发射光谱法(ICP-OES),分析测定了青藏高原地区产藏雪莲水母雪兔子中的矿质元素.结果显示,藏雪莲水母雪兔子中含有K、Ca、P、Mg、Al、S、Fe、Si、Na、Mn、Ti、Sr、Ba、B、Zn、Cu、Cr等17种矿质元素,定量分析结果相对标准偏差...     

新型气升式反应器中浸没/非浸没周期对水母雪莲细胞培养的影响

应用新型周期浸没气升式反应器，进行了水母雪莲细胞的浸没悬浮和非浸没静置交替循环培养，研究了培养过程中的关键条件?浸没/非浸没周期对水母雪莲细胞生长、黄酮合成及细胞聚集体调控的影响，发现5 min/4 h为最佳的循环周期. 在此循环周期下，当通气量为40 L/h、接种量(干重DW)为2~3 g/L时，经20 d的培养，细胞聚集体分布最佳, 细胞干重达9.6 g/L，黄酮类化合物的含量及黄酮类化合物的产量分别可达35.2 mg/g (DW)及338 mg/L.     

Three Pairs of Novel Enantiomeric 8-O-4′ Type Neolignans from Saussurea medusa and Their Anti-inflammatory Effects In Vitro

Three pairs of novel enantiomeric 8-O-4′ type neolignans (1a/1b–3a/3b), together with seven known analogues (4–10), were isolated from the whole plants of Saussurea medusa. Their structures were elucidated by extensive spectroscopic data analysis and electric circular dichroism (ECD) calculations after chiral separations. All compounds were obtained from S. medusa for the first time, and compounds 1–3 and 5–10 had never been obtained from the genus Saussurea previously. The anti-inflammatory activities of the compounds were evaluated by determining their inhibitory activities on the production of NO and inducible nitric oxide synthase (iNOS) expression in LPS-stimulated RAW 264.7 cells. Compounds (+)-1a, (−)-1b and 5–7 inhibited NO production and had IC₅₀ values ranging from 14.3 ± 1.6 to 41.4 ± 3.1 μM. Compound 7 induced a dose-dependent reduction in the expression of iNOS in LPS-treated RAW 264.7 cells. Molecular docking experiments showed that all active compounds exhibited excellent docking scores (<−7.0 kcal/mol) with iNOS. Therefore, compounds (+)-1a, (−)-1b and 5–7 isolated from the whole plants of S. medusa may have therapeutic potential in inflammatory diseases.     

光温条件对新疆雪莲愈伤组织反应器培养后再分化能力的影响

研究了光质、光周期、昼夜温差和低温处理时间对反应器大规模培养后的新疆雪莲愈伤组织再分化能力的影响. 实验结果表明,长波长光更有利于愈伤组织的再分化. 光照时间显著影响愈伤组织的再分化能力,光照时间为16 h/d时愈伤组织的分化频率和平均出芽数达到最大,分别为76.7%和1.8个/块. 昼夜温差与愈伤组织的再分化能力呈显著负相关. 与水母雪莲不同,低温处理并不利于新疆雪莲愈伤组织的再分化. 新疆雪莲愈伤组织的再分化能力与同工酶的种类和活性密切相关.     

原子荧光光谱法测定绵头雪莲花中的痕量砷

采用原子荧光光谱法测定绵头雪莲花中的微量砷,测量结果为0.5994 mg/kg,方法检出限为0.02μg/L,回收率为93%~105%,RSD为0.607%。测量结果表明绵头雪莲花中的砷含量没有超出国家安全标准,而且原子荧光光谱法对于测定绵头雪莲花中的砷含量具有精密度高、操作简单、快速和灵敏度高的优点。     

Quantitative and Functional Assessment of the Influence of Routinely Used Cryopreservation Media on Mononuclear Leukocytes for Medical Research

Quantitative and functional analysis of mononuclear leukocyte populations is an invaluable tool to understand the role of the immune system in the pathogenesis of a disease. Cryopreservation of mononuclear cells (MNCs) is routinely used to guarantee similar experimental conditions. Immune cells react differently to cryopreservation, and populations and functions of immune cells change during the process of freeze–thawing. To allow for a setup that preserves cell number and function optimally, we tested four different cryopreservation media. MNCs from 15 human individuals were analyzed. Before freezing and after thawing, the distribution of leukocytes was quantified by flow cytometry. Cultured cells were stimulated using lipopolysaccharide, and their immune response was quantified by flow cytometry, quantitative polymerase chain reaction (qPCR), and enzyme-linked immunosorbent assay (ELISA). Ultimately, the performance of the cryopreservation media was ranked. Cell recovery and viability were different between the media. Cryopreservation led to changes in the relative number of monocytes, T cells, B cells, and their subsets. The inflammatory response of MNCs was altered by cryopreservation, enhancing the basal production of inflammatory cytokines. Different cryopreservation media induce biases, which needs to be considered when designing a study relying on cryopreservation. Here, we provide an overview of four different cryopreservation media for choosing the optimal medium for a specific task.     

Cryopreservation of Gametes and Embryos and Their Molecular Changes

The process of freezing cells or tissues and depositing them in liquid nitrogen at –196 °C is called cryopreservation. Sub-zero temperature is not a physiological condition for cells and water ice crystals represent the main problem since they induce cell death, principally in large cells like oocytes, which have a meiotic spindle that degenerates during this process. Significantly, cryopreservation represents an option for fertility preservation in patients who develop gonadal failure for any condition and those who want to freeze their germ cells for later use. The possibility of freezing sperm, oocytes, and embryos has been available for a long time, and in 1983 the first birth with thawed oocytes was achieved. From the mid-2000s forward, the use of egg vitrification through intracytoplasmic sperm injection has improved pregnancy rates. Births using assisted reproductive technologies (ART) have some adverse conditions and events. These risks could be associated with ART procedures or related to infertility. Cryopreservation generates changes in the epigenome of gametes and embryos, given that ART occurs when the epigenome is most vulnerable. Furthermore, cryoprotective agents induce alterations in the integrity of germ cells and embryos. Notably, cryopreservation extensively affects cell viability, generates proteomic profile changes, compromises crucial cellular functions, and alters sperm motility. This technique has been widely employed since the 1980s and there is a lack of knowledge about molecular changes. The emerging view is that molecular changes are associated with cryopreservation, affecting metabolism, cytoarchitecture, calcium homeostasis, epigenetic state, and cell survival, which compromise the fertilization in ART.