Lycium ruthenicum Murr. treatment attenuates APP_(SWE)/PS1ΔE9 mouse model-like mitochondrial dysfunction in Slc25a46 knockout mouse model

Mitochondrial dysfunction is proposed to be substantially associated with ageing and ageing-related diseases like Alzheimer's disease(AD). However, it is unclear whether different mouse models with mitochondrialrelated diseases have similar changes in mitochondrial morphology of the same tissues. Moreover, whether similarities in mitochondrial morphology can be a suitable marker for screening and/or discovering mitochondrial-protective substances remains unknown. Mitochondria morphology in different tissues of a novel mitochondrial outer membrane protein Slc25a46 knockout mouse and a traditional APP_(SWE)/PS1ΔE9 transgenic mouse were examined using transmission electron microscope(TEM). Both young Slc25a46 knockout mice and aged APP_(SWE)/PS1ΔE9 mice models showed similar mitochondrial damage in cerebellum tissues. The results indicated that different mitochondrial-related diseases shared similar alteration and defects in mitochondrial morphology. Furthermore, Lycium ruthenicum Murr. extract, a bioactive food substance with cognition-improving property, could effectively improve muscle strength and increase body weight in the Slc25a46 knockout mice. These findings suggest that mitochondrial morphology defects in mice models, particularly in the mitochondrial compartment, represent a unified and effective marker for screening and validating natural product-derived functional substances with mitochondrial protective properties. It also holds potential application in mitochondrial-impaired senile neurodegenerative diseases, especially in AD.

黑果枸杞香气成分和多酚类物质的组成研究

随着人类对健康和养生重视程度的不断提高,具有良好功效的药食同源植物黑果枸杞受到越来越多的关注,研究黑果枸杞中具有药理活性物质的化学成分,对其深加工和进一步开发利用具有重要的意义。该文对甘肃酒泉产的黑果枸杞鲜果的香气成分及多酚类物质的化学组成进行分析,采用顶空固相微萃取-气相色谱-质谱联用技术定性定量分析黑果枸杞鲜果香气成分;采用超高效液相色谱-电喷雾四级杆串联飞行时间质谱技术定性分析黑果枸杞鲜果中多酚类物质的组成。从黑果枸杞香气成分中鉴定出醇类、醛酮类、酯类、烃类等挥发性化学成分63种,占香气成分总量的95.4%,其中25种化合物为我国国家标准GB 2760—2014《食品安全国家标准食品添加剂使用标准》规定的食品用合成香料;香气成分中醇类化合物共计26种,总含量占比最高(44.98%)。黑果枸杞多酚经提取和HPD-500大孔吸附树脂纯化后进行液相色谱-质谱联用测试,定性鉴别出73种多酚化合物,其主要由黄酮类和酚酸类等化合物组成。该研究初步确定了黑果枸杞香气和多酚类物质的化学成分,为黑果枸杞药用及食用功效的深入研究奠定了基础。

黑果枸杞和黄果枸杞的广泛靶向代谢组学分析

目的:全面分析和鉴定枸杞果实中的代谢物,比较不同品种间代谢物的差异,为深入挖掘和利用不同品种枸杞提供参考。方法:以黑果枸杞和黄果枸杞干果为材料,利用超高效液相色谱-串联质谱进行广泛靶向代谢组学分析,鉴定和比较两种枸杞的代谢物。结果:两种枸杞果实中共检测出1098种代谢产物,包括氨基酸及其衍生物、酚酸、黄酮、生物碱、脂质等12类,其中酚酸、黄酮和生物碱3类代谢产物占比最高。通过主成分分析和聚类分析发现,黄果枸杞与黑果枸杞相比,存在235种差异代谢物上调和252种差异代谢物下调。京都基因与基因组百科全书富集分析发现,不同品种枸杞间的差异代谢物主要涉及黄酮生物合成,托烷、哌啶和吡啶生物碱的生物合成,次生代谢物的生物合成,ABC转运等途径。结论:不同品种对枸杞代谢物的含量影响较大,本研究结果对黑枸杞和黄枸杞的开发和利用提供一定的理论指导。

黑果枸杞多糖结构表征及对乙醇诱导胃黏膜上皮细胞损伤的影响

采用热水浸提法提取黑果枸杞多糖(Lycium ruthenicum Murr. polysaccharide,LRMP),解析其单糖组成、糖苷键连接方式、链构象等结构参数,并评价其对乙醇诱导胃黏膜上皮细胞(gastric mucosal epithelial cells,GES-1)损伤的保护效果。结果表明,LRMP由阿拉伯糖、木糖、半乳糖、葡萄糖、岩藻糖和半乳糖醛酸组成,结构末端主要由阿拉伯糖、木糖、葡萄糖和半乳糖残基组成,在溶液中呈典型的无规卷曲构象,均方半径为(80.4±1.0)nm。LRMP对GES-1细胞无毒性作用,质量浓度600μg/mL LRMP可以显著抑制乙醇对GES-1细胞造成的损伤,同时恢复细胞内超氧化物歧化酶的活性,降低GES-1细胞凋亡率。综上所述,LRMP作为功能性多糖在健康食品开发中具有潜力。

黑果枸杞不同发育时期果实花色苷合成的转录组分析

【目的】探究黑果枸杞果实发育过程中花色苷合成的分子机制,对深入理解黑果枸杞花色苷合成调控网络具有重要意义。【方法】选取青果期、转色初期、转色后期、成熟期和完全成熟期的黑果枸杞果实进行转录组测序,挖掘与黑果枸杞果实花色苷合成相关的候选基因。【结果】随黑果枸杞果实发育,花色苷含量逐渐升高,成熟期及完全成熟期果实中的花色苷含量显著高于青果期、转色初期及转色后期。5个发育时期共筛选出13540个差异表达基因(differentially expressed genes,DEGs),以青果期为对照,随果实发育,各阶段DEGs数量逐渐增多。GO分析发现,DEGs共同富集的子集有苯丙烷代谢过程、多糖分解代谢过程、苯丙烷生物合成过程、类黄酮生物合成过程、细胞壁大分子代谢过程、膜锚定成分和营养库活性通路。KEGG分析表明,DEGs显著富集于类黄酮生物合成、苯丙烷生物合成、植物激素信号转导、二苯乙烯类、二芳基庚烷类和姜辣素生物合成及角质和木栓质和蜡质的生物合成通路。分析DEGs表达量与花色苷含量相关性,筛选出参与黑果枸杞果实发育过程花色苷合成的候选基因36个,主要包括10个花色苷合成通路结构基因,4个转录因子,9个ABA、8个GA及5个JA信号转导通路基因。RT-qPCR验证其中10个基因的表达趋势,与转录组数据一致。【结论】黑果枸杞果实发育过程中,花色苷合成途径的结构基因、转录因子及ABA、GA和JA信号转导通路中基因的表达调控果实着色。

Effects of NaCl Stress on Seed Germination of Lycium ruthenicum Murr.

[Objective]To study the seeds germination of Lycium ruthenicum Murr.under different concentrations of NaCl,as well as to find the optimal concentration of NaCl for the germination of L.ruthenicum.[Method]The seeds of L.ruthenicum were treated with different concentrations of NaCl,and the state of seed germination was measured.[Result]With the increasing of concentration of NaCl,the seed germination rate of L.ruthenicum showed an obvious increasing trend.when the concentration was of 0.3%-0.4 %,the germination rate was the highest,and when the concentration of NaCl was greater than 0.4,the germination rate showed a decline trend.[Conclusion]After treated with appropriate concentrations of NaCl before sowing,the germination rate of seeds of L.ruthenicum would increase.

Karyotype Analysis of Lycium ruthenicum Murr.

Karyotype analysis of Lycium ruthenicum Murr. was carried out in this study. The results showed that the chromosome number was 2n=2x=24; the arm index was 48; the ratio of the longest chromosome to the shortest one was 1.31; the proportions of chromosomes with arm ratio higher than 2 was 0.08; the asymmetry index was 57.02; the karyotype type was 2A; and the karyotype formula was 2n=-24=20m＋4sm.

食品加工条件对黑枸杞花青素稳定性的影响

黑枸杞富含花青素,花青素具有多种生理活性,然而其稳定性较差。文章研究了不同食品加工条件对黑枸杞花青素稳定性的影响,包括温度、pH值、光照、食品添加剂和基质等。结果表明,在20℃、pH3、避光条件下的黑枸杞花青素相对稳定,保存率分别为(89.74±5.54)%、(69.56±1.44)%、(57.85±0.50)%。H_(2)O_(2)、Na_(2)SO_(3)、苯甲酸钠和3种糖(葡萄糖、蔗糖、乳糖)均会大幅降低花青素的热稳定性(65℃,30 min)。除抗坏血酸外,草酸、酒石酸和柠檬酸增强花青素热稳定性的效果依次增大。在40℃加速贮存7 d后,草酸对花青素的稳定性仍然具有较好的效果。总之,黑枸杞花青素在食品加工过程中最好在低温、酸性和避光条件下贮存,而在选用食品基质和添加剂时也应该注意其对花青素稳定性的影响。该研究结果为黑枸杞花青素类食品的加工提供了理论依据。

黑枸杞花青素联合人脂肪源性血管外膜细胞支持脐血造血干/祖细胞的增殖

背景:黑枸杞花青素(Anthocyanins in Lycium ruthenicum Murr,ALRM)是黑枸杞中重要的活性成分之一,具有抗氧化、免疫调节等功效。人脂肪源性血管外膜细胞(CD146^(+)hAD-PCs)是骨髓间充质干细胞的前体细胞,在体外具有促进造血干/祖细胞增殖与分化的功能。ALRM联合CD146^(+)hAD-PCs对脐血造血干/组细胞的体外支持作用有待于研究。目的:探讨ALRM联合CD146^(+)hAD-PCs对脐血CD34^(+)造血干/祖细胞体外扩增的支持作用。方法:CCK-8法检测不同质量浓度ALRM(0,200,400,600,800,1000 mg/L)对CD146^(+)hAD-PCs增殖的影响;流式细胞术检测ALRM对CD146^(+)hAD-PCs细胞周期的影响。共培养实验分为空白组、ALRM组、CD146^(+)hAD-PCs组、ALRM+CD146^(+)hAD-PCs组,分析ALRM联合CD146^(+)hAD-PCs对脐血CD34^(+)造血干/祖细胞的体外支持作用。共培养1,2,4周,比较扩增后细胞数量、集落形成单位数量,流式细胞仪检测细胞免疫表型,ELISA检测细胞因子水平。结果与结论:(1)ALRM质量浓度为200 mg/L时,CD146^(+)hAD-PCs活力最高,CD146^(+)hAD-PCs的G_(0)/G_(1)期细胞比例下降,S期、G_(2)/M期细胞比例上升(P<0.01)。(2)脐血CD34^(+)造血干/祖细胞数量变化:在共培养1,2,4周时ALRM+CD146^(+)hAD-PCs组高于ALRM组(P均<0.05),在共培养2,4周时ALRM+CD146^(+)hAD-PCs组高于CD146^(+)hAD-PCs组(P均<0.05),ALRM组与空白组随着共培养时间延长细胞数量逐渐减少。(3)集落形成能力及免疫表型分析:在共培养1,2周时ALRM+CD146^(+)hAD-PCs组的集落形成单位数量高于CD146^(+)hAD-PCs组和ALRM组(P均<0.05);在共培养1,2,4周时ALRM+CD146^(+)hAD-PCs组CD45^(+)、CD34^(+)CD33^(-)细胞比例高于CD146^(+)hAD-PCs组(P均<0.01)。(4)细胞因子变化:在共培养4周时ALRM+CD146^(+)hAD-PCs组的白细胞介素2水平高于ALRM组、CD146^(+)hAD-PCs组(P<0.05);在共培养2,4周时ALRM+CD146^(+)hAD-PCs组白细胞介素3水平高于CD146^(+)hAD-PCs组(P<0.05);在共培养1周时ALRM+CD146^(+)h AD-PCs组的粒细胞集落刺激因子水平高于CD146^(+)hAD-PCs组,在共培养2周时高于ALRM组、CD146^(+)hAD-PCs组(P<0.01);在共培养1,2,4周时ALRM组、ALRM+CD146^(+)hAD-PCs组的干扰素γ水平低于CD146^(+)hAD-PCs组(P<0.05)。(5)由于空白组无基质细胞,脐血CD34^(+)造血干/祖细胞在共培养1周之后就无法计数,未进行免疫表型、集落分析和细胞因子检测。(6)结果表明:ALRM可以通过促进CD146^(+)hAD-PCs增殖和细胞周期转化进而促进脐血CD34^(+)造血干/祖细胞的体外扩增,在造血干细胞移植研究方面具有重要价值。

黑果枸杞‐酿酒葡萄混合果酒酿造过程中花色苷稳定性研究

针对黑果枸杞加工过程中存在的花色苷不稳定的关键瓶颈问题,以黑果枸杞鲜果为原料,辅以酿酒葡萄共发酵酿制果酒。分析不同酿造因子对果酒多酚、花色苷及色泽的影响;明确影响酒体中花色苷含量及稳定性的主要因子,通过正交优化试验得到最佳酒精发酵工艺。结果表明:黑果枸杞与酿酒葡萄质量比以及pH值对果酒花色苷含量和色泽影响程度较大,但对多酚含量影响作用较小,而酵母菌类型对黑果枸杞果酒多酚、花色苷和色泽影响均较小;复配酿酒葡萄可减少花色苷损失,提高黑果枸杞果酒中花色苷的稳定性。最佳酒精发酵工艺条件为黑果枸杞与酿酒葡萄质量比1∶4、果胶酶添加量0.70g/L、酿酒酵母添加量0.25g/L,发酵后酰化花色苷[矮牵牛素-3-O-芸香糖(反式-p-香豆酰基)-5-O-葡萄糖苷]含量可达(459.51±3.66)mg/L。

黑果枸杞MSAP技术体系的构建

为利用甲基化敏感多态性扩增技术(MSAP)构建黑果枸杞MSAP技术反应体系,本试验采用CTAB法提取黑果枸杞DNA,通过双酶切、预扩增和选择性扩增对16对引物的多态性进行初步筛选并构建MSAP技术体系。结果表明:双酶切进行12 h,DNA模板能够酶切完全;预扩增反应产物条带集中在250~1000 bp处,选择性扩增可筛选出6对引物组合,条带明显。本研究为后续黑果枸杞DNA甲基化水平的相关分析研究奠定了基础。

松杉灵芝孢子油超声波辅助提取响应面优化

以松杉灵芝孢子粉为原料,提取灵芝孢子油。通过单因素试验研究超声功率、超声时间、超声温度对灵芝孢子油得率的影响,并利用响应面法对松杉灵芝孢子油超声波辅助提取工艺进行优化。结果表明,超声功率300 W,超声时间30 min,超声温度25℃,在此工艺条件下灵芝孢子油的得率为28.5%。

液态深层发酵松杉灵芝提取麦角硫因的技术研究

本文通过单因素以及正交试验确定液态深层发酵松杉灵芝提取麦角硫因的提取条件。结果表明,最佳提取条件为最大允许装液量的60%,转速200 r·min^(-1),发酵液pH=6,发酵温度30℃,发酵通气量0.6 L·min^(-1)。

黑果枸杞果实中主要成分的傅里叶变换近红外光谱预测模型构建

使用傅里叶变换近红外光谱(FT-NIR)结合化学计量学方法,开发了一种黑果枸杞干果和鲜果中主要成分(总糖、还原糖、总酸、氨态氮、花青素、原花青素、总酚、黄酮和多糖)含量的预测方法。首先比较了11种原始光谱的预处理方式,筛选出每种成分的最优预处理方法。然后比较了利用偏最小二乘(PLS)、区间偏最小二乘(iPLS)和联合区间偏最小二乘(siPLS)算法建立的模型,最终确定采用siPLS建模。结果表明:总酸、氨态氮、花青素、原花青素、总酚和黄酮的交叉验证相关系数(Rc)和预测集相关系数(R_(P))均大于0.9818,相对分析误差(RPD)均大于2.5,模型效果优异,总糖、还原糖和多糖的建模效果良好,建立的定标模型均可以用于实际检测。验证集样本实测值与预测值无显著性差异,预测误差在±0.1%,模型的预测结果可信度高。本研究建立的预测模型,可以实现黑果枸杞干果和鲜果中主要成分含量的无损、快速、准确检测。

黑果枸杞中原花青素的提取及其稳定性研究

以黑果枸杞中原花青素为研究对象,通过单因素实验分别考查乙醇浓度、料液比、提取时间、提取温度对原花青素提取率的影响,同时通过响应面法优化提取实验,得到提取最优工艺。另外,分别考查了不同种类、不同浓度的糊精和氨基酸对原花青素稳定性的影响,结果表明,其对原花青素具有一定的稳定性作用。

松干基褐腐病菌Phellinus weirii (Murr.) Gilgertson的风险管理浅析

本文通过对松干基褐腐病病原菌的地理分布、危害的寄主、病原物形态特征、生物学特性、危害情况、进入扩散可能性、经济重要性等进行风险分析,评价松干基褐腐病病原菌的风险性,并提出了检疫管理措施。

Study on Tissue Culture and Rapid Propagation of Lycium ruthenicum Murr.

[Objective] This study aimed to establish a tissue culture and rapid propagation system for Lycium ruthenicum Murr.[Method] Using tender stems of L.ruthenicum as explants,MS as basic culture medium,the effects of different factors on primary culture,subculture and rooting of L.ruthenicum plantlets were investigated.[Result] The most appropriate medium for primary culture of L.ruthenicum was MS ＋ ZT 0.2 mg/L ＋ IBA 0.01 mg/L,in which axillary buds grew well and were rarely vitrified with the germination rate of 88.73%.In addition,ZT exerted significantly better effects on subculture and proliferation of L.ruthenicum plantlets than 6-BA.The most appropriate medium for subculture and proliferation of L.ruthenicum plantlets was MS ＋ ZT 0.15 mg/L ＋ IBA 0.01 mg/L,in which L.ruthenicum plantlets grew rapidly and robustly without vitrification,and the proliferation multiple reached 5.83 times.The most appropriate medium for rooting of L.ruthenicum plantlets was MS ＋ IBA 1.0 mg/L,in which the rooting rate reached 100%.The most appropriate substrate for transplanting and hardening of L.ruthenicum plantlets was humus soil：perlite = 1：1,in which L.ruthenicum plantlets grew well with the survival rate of 92.37%.[Conclusion] This study provided theoretical basis for largescale production and popularization of L.ruthenicum.

Copper toxicosis gene MURRl is not changed in Wilson disease patients with normal blood ceruloplasmin levels

AIM： To analyze our Wilson disease patient cohort （n = 106） for alterations in the gene coding for MURR1. METHODS： Patients with an established diagnosis of Wilson disease but normal ceruloplasmin blood levels were chosen for our study （n = 14）. Patients with two known disease-causing mutations in the ATPTB gene were not included. The three exons of the human MURR1 gene were sequenced after amplification of the genomic DNA by polymerase chain reaction. RESULTS： Our study did not reveal any mutations leading to an amino acid change in the MURR1 sequence of Wilson disease patients. A polymorphism at 472 bp of the coding sequence could be confirmed. CONCLUSION： The MURRI gene plays no role in the pathogenesis of Wilson disease patients with normal serum ceruloplasmin levels.

Neuroprotective effect of Spilanthes acmella Murr. on pesticide-induced neuronal cells death

Objective: To investigate protective effects of Spilanthes acmella(S,acmella) Murr,extracts against pesticide-induced neuronal cells death and to elucidate the underlying molecular mechanism in dopaminergic(SH-SY5Y) cells lines,Methods: Cell viability of SH-SY5 Y cells was studied by treating the cells with various concentration of pirimicarb for 24 hr,Neuroprotective effect of S,acmella Murr,extracts was investigated by adding the plant extracts to the medium for 24 hr prior to the incubation with 100 μM H_2O_2 or with pirimicarb for 24 hr,Control-untreated cells were incubated with the culture medium,Cell viability was measured by MTT assay,calpain and calpastatin expressions were analyzed by Western blotting and immunocytochemistry,Results: Pretreatment of SH-SY5 Y cells with S,acmella Murr,extracts(1 μg/m L) for 24 hr significantly increased the dopaminergic neurons in pirimicarb-induced neurotoxicity,In addition,pretreatment with the S,acmella Murr,extracts led to decreased calpain but increased calpastatin protein levels,Conclusion: S,acmella Murr,extracts exerted neuroprotective effect,via an alteration of calcium homeostasis,against pirimicarb induced neurotoxicity,The S,acmella Murr,might be a potential natural candidate with neuroprotective activity.

Bidirectional solid fermentation using Trametes robiniophila Murr. For enhancing efficacy and reducing toxicity of rhubarb (Rheum palmatum L.)

Objective:To investigate the potential of bidirectional solid fermentation of rhubarb (Rheum palmatum L.) for reducing its toxicity and enhancing its medicinal efficacy.Methods:The fungus Trametes robiniophila Murr.was inoculated into rhubarb.The chemical ingredients as well as antioxidant,antibacterial,and anticancer activities of fermented and unfermented rhubarb extracts were then determined.Results:After fermentation,levels of anthraquinone glycosides (purgative ingredients) decreased significantly,while the level of anthraquinone aglycone increased.The level of gallic acid was also reduced after fermentation.Ethanol extract of rhubarb (0.8 mg/mL) exhibited DPPH-scavenging activity of 7.6% ± 0.8% while the blank control (0.8 mg/mL rhubarb)showed 31.3% ± 2.0% activity.Antibacterial activities in fermented samples were found to be enhanced compared with unfermented samples and anticancer activity was evident at concentrations of 2000 and 5000 μg/mL.Conclusion:Bidirectional solid fermentation appears to be an effective processing method that can be used to improve the efficacy and reduce the toxicity of rhubarb.