Cloning and Characterization of a Lycium chinense Carotenoid Isomerase Gene Enhancing Carotenoid Accumulation in Transgenic Tobacco

Carotenoid isomerase （CRTISO）is a key enzyme that catalyzes the conversion of cis-lycopene to all- trans tycopene. In this study, we isolated and characterized the CRTISO gene from Lycium chinense （LcCRTISO） for the first time. The open reading flame of LcCRTISO was 1 815 bp encoding a protein of 604 amino acids with a molecular mass of 66.24 kDa. Amino acid sequence analysis revealed that the LcCRTISO had a high level of simi- larity to other CRTISO. Phylogenetic analysis displayed that LcCRTISO kept a closer relationship with the CRTISO of plants than with those of other species. Semi-quantitative PCR analysis indicated that LcCRTISO gene was expressed in all tissues tested with the highest expression in maturing fruits. The overexpression of LcCRTISO gene in transgenic tobacco resulted in an increase of total carotenoids in the leaves with [3-carotene and lutein being the predominants. The results obtained here clearly suggested that the LcCRTISO gene was a promising candidate for carotenoid production.

Drought stress,mercuric chloride,andβ-mercaptoethanol effects on hydraulic characteristics of three cultivars of wolfberry(Lycium chinense)

In order to further understand the effects of drought stress on hydraulic characteristics and the relationship between hydraulic conductivity and aquaporins or water channels of root systems of three wolfberry cultivars(Lycium chinense Mill.),hydraulic conductivity of 2-yearold pot-grown seedlings was measured under drought stress,rewatering,and treatment with exogenous mercuric chloride andβ-mercaptoethanol.Under moderate and severe drought stress levels,the most significant decrease of hydraulic conductivity was 37.3%and 24.0%,respectively,in the‘Ningqi 5’cultivar compared with the nonstressed control.After rewatering,the rate of recovery in specific conductivity was most rapid in the‘Mengqi 1’cultivar,at 0.058 and 0.072 kg MPa^-1 m^-2 s^-1 h^-1 under moderate and severe drought stress levels,respectively.The‘Mengqi 1’cultivar had the highest recovery degree of hydraulic conductivity under two concentrations ofβ-ME(500 or 1000 lmol L^-1),reaching 82.4%and 88.5%,respectively,of the initial conductivity.The adaptive capacity of hydraulic conductivity in the‘Ningqi 5’cultivar was weaker than in the‘Ningqi 1’and‘Mengqi 1’cultivars under drought stress.The recovery capacity of hydraulic conductivity in‘Mengqi 1’cultivar was stronger than the‘Ningqi 1’and‘Ningqi 5’cultivars after rewatering.Aquaporins of the‘Ningqi 1’cultivar root systems had the highest binding affinity with mercuric chloride,which was the most likely cause in the decrease in hydraulic conductivity,whereas aquaporins of‘Mengqi 1’root systems had the weakest binding affinity.The inhibitory effect of mercuric chloride was readily eliminated byβ-mercaptoethanol in the‘Mengqi 1’cultivar.The hydraulic characteristics of this cultivar were more sensitive to drought,mercuric chloride andβ-mercaptoethanol than the other cultivars.

Effects of Lycium barbarum polysaccharide on cytokines in adolescents with subthreshold depression:a randomized controlled study

Strong evidence has accumulated to show a correlation between depression symptoms and inflammatory responses.Moreover,anti-inflammatory treatment has shown partial effectiveness in alleviating depression symptoms.Lycium barbarum polysaccharide(LBP),derived from Goji berries,exhibits notable antioxidative and anti-inflammatory properties.In our recent double-blinded randomized placebo-controlled trial,we found that LBP significantly reduced depressive symptoms in adolescents with subthreshold depression.It is presumed that the antidepressant effect of LBP may be associated with its influence on inflammatory cytokines.In the double-blinded randomized controlled trial,we enrolled 29 adolescents with subthreshold depression and randomly divided them into an LBP group and a placebo group.In the LBP group,adolescents were given 300 mg/d LBP.A 6-week follow up was completed by 24 adolescents,comprising 14 adolescents from the LBP group(15.36±2.06 years,3 men and 11 women)and 10 adolescents from the placebo group(14.9±1.6 years,2 men and 8 women).Our results showed that after 6 weeks of treatment,the interleukin-17A level in the LBP group was lower than that in the placebo group.Network analysis showed that LBP reduced the correlations and connectivity between inflammatory factors,which were associated with the improvement in depressive symptoms.These findings suggest that 6-week administration of LBP suppresses the immune response by reducing interleukin-17A level,thereby exerting an antidepressant effect.

不同浓缩方式对枸杞汁品质特性的影响

枸杞汁富含多种营养和保健功能成分,浓缩汁是其重要的流通形式。为探究浓缩方式对枸杞汁品质特性的影响,本文对比了热浓缩(heating concentration,HC)、真空浓缩(vacuum concentration,VC)、二级冷冻浓缩(secondary freezing concentration,SFC)、热联合一级冷冻浓缩(heating-assisted primary freezing concentration,HAPFC)、真空联合一级冷冻浓缩(vacuum-assisted primary freezing concentration,VAPFC)五种浓缩方法,考察浓缩前后枸杞汁色泽、还原糖、糖组分、有机酸组分、总酚、总黄酮、类胡萝卜素、DPPH及FRAP等的变化。结果表明,真空浓缩对色泽的保护效果显著优于其他处理组,能较好地保留枸杞汁中的还原糖,对苹果酸和酒石酸影响较小。真空浓缩能较好地保留枸杞汁中的总酚、总黄酮含量,较其他处理组分别高8.76%~21.40%和5.41%~13.77%;真空浓缩能有效保留枸杞汁中的类胡萝卜素,且DPPH自由基清除能力和铁离子还原能力较其他处理组高6.98%~29.77%和5.97%~26.76%。因此,真空浓缩对枸杞浓缩汁品质的保护效果最好,是一种适宜枸杞汁浓缩的方式。

Efficacy of Lycium barbarum polysaccharide in adolescents with subthreshold depression: interim analysis of a randomized controlled study

Subthreshold depression is a highly prevalent condition in adolescents who are at high risk for developing major depressive disorder.In preclinical models of neurological and psychiatric diseases,Lycium barbarum polysaccharide(LBP)extracted from Goji berries had antidepressant effects including but not limited to anti-oxidative and anti-inflammatory properties.However,the effect of LBP on subthreshold depression is unclear.To investigate the clinical efficacy and safety of LBP for treating subthreshold depression in adolescents,we conducted a randomized,double-blind,placebo-controlled trial(RCT)with 29 adolescents with subthreshold depression recruited at The Fifth Affiliated Hospital of Guangzhou Medical University.The participants were randomly assigned to groups where they received either 300 mg LBP(LBP group,n=15,3 boys and 12 girls aged 15.13±2.17 years)or a placebo(placebo group,n=14,2 boys and 12 girls aged 15±1.71 years)for 6 successive weeks.Interim analyses revealed that the LBP group exhibited a greater change in Hamilton Depression Scale(HAMD-24)scores relative to the baseline and a higher remission rate(HAMD-24 total score≤7)at 6 weeks compared with the placebo group.Scores on the Beck Depression Inventory-II(BDI-II),Pittsburgh Sleep Quality Index(PSQI),Kessler Psychological Distress Scale(Kessler),and Screen for Child Anxiety-Related Emotional Disorders(SCARED)were similar between the LBP and placebo groups.No side effects related to the intervention were reported in either group.These results indicate that LBP administration reduced depressive symptoms in adolescents with subthreshold depression.Furthermore,LBP was well tolerated with no treatment-limiting adverse events.Clinical trials involving a larger sample size are needed to further confirm the anti-depressive effects of LBP in adolescents with subthreshold depression.This study was approved by the Medical Ethics Committee of the Fifth Affiliated Hospital of Guangzhou Medical University(Guangzhou,China;approval No.L2019-08)on April 4,2019 and was registered on ClinicalTrials.gov(identifier:NCT04032795)on July 25,2019.

不同盐胁迫条件下枸杞种子的耐锂特性研究

以3个枸杞品种(宁杞1号、宁杞5号、宁杞7号)为试材,研究不同浓度NaCl溶液对枸杞种子萌发的影响,以及盐胁迫下不同浓度LiCl溶液处理对3个品种枸杞发芽率、发芽势、发芽指数、根长、芽长和活力指数的影响。结果表明,随着盐浓度的不断升高,对宁杞1号的根长和芽长均起抑制作用;对宁杞5号的根长和芽长则是先促进后抑制;而对宁杞7号的根长表现出抑制作用,对芽长表现出先促进后抑制。低锂浓度对宁杞1号、宁杞5号和宁杞7号的发芽率、发芽势、发芽指数、根长、芽长和活力指数起促进作用,其中宁杞1号较宁杞5号和宁杞7号具有更强的耐锂性。

北方枸杞化学成分及其α-葡萄糖苷酶抑制活性研究

目的:研究北方枸杞Lycium chinense var.potaninii果实的化学成分及其α-葡萄糖苷酶抑制活性。方法:运用大孔吸附树脂、硅胶、ODS反相硅胶以及制备高效液相等技术对北方枸杞干燥果实的95%乙醇提取物进行分离纯化,根据波谱学方法鉴定化合物结构,并对化合物进行α-葡萄糖苷酶抑制活性筛选。结果:从北方枸杞果实的乙酸乙酯部位中分离得到16个化合物,分别鉴定为:N-顺式阿魏酰基酪胺(1)、methyl-2-[2-formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl]-3-(4-hydroxyphenyl)propanoate(2)、菜豆酸(3)、2E,4Z-脱落酸(4)、催吐萝芙木醇(5)、(E)-3-(3,4,5-trimethoxyphenyl)acrylic acid(6)、阿魏酸甲酯(7)、阿魏酸(8)、3-甲氧基-4-羟基苯丙酸(9)、东莨菪内酯(10)、5-羟基麦芽酚(11)、对羟基苯乙酮(12)、反式对羟基肉桂酸(13)、香草乙酮(14)、香草醛(15)、β-谷甾醇(16)。结论:其中,化合物3~5为首次从枸杞属植物中分离得到,化合物1、2、6~16为首次从该植物中分离得到。化合物1对α-葡萄糖苷酶具有显著的抑制作用,其IC50为(24.25±1.12)μmol/L。

超声波辅助双水相法提取枸杞叶黄酮及其胰脂肪酶抑制活性

[目的]开发一种安全、无毒且高效的枸杞叶黄酮类化合物提取方法。[方法]采用超声波辅助双水相法实现高得率、绿色的枸杞叶黄酮类化合物提取,并评价产物活性。通过单因素和Plackett-Burman(PB)试验考察各因素对黄酮提取效果的影响,选取聚乙二醇(PEG400)使用量、硫酸铵使用量、料液比3个因素通过响应面中心组合试验设计优化提取条件;通过胰脂肪酶活性抑制试验评价提取物活性。[结果]超声波辅助双水相法提取枸杞叶黄酮最佳工艺条件为:聚乙二醇PEG相对分子质量400,PEG质量分数28%、硫酸铵质量分数16%、超声功率250 W,超声处理时间40 min、超声处理温度40℃、料液比1∶30 (g/g),该条件下所提取的枸杞叶黄酮提取物的总黄酮含量为(486.30±3.29) mg/g,其对胰脂肪酶活性具有显著抑制作用,半数抑制质量浓度(IC50)为1.06 mg/mL。[结论]最优提取条件下,超声波辅助双水相法所提取的枸杞叶黄酮得率高、溶剂残留低,该方法所得枸杞叶黄酮对胰脂肪酶具有良好的抑制作用。

基于SSR荧光标记的121份野生枸杞种质遗传结构解析

以采集的121份野生枸杞(Lycium barbarum L.)种质为材料,用9对SSR引物进行扩增,采用GeneALEX、Power Marker软件进行多态性分析,利用NTSYS、STRUC-TURE软件进行遗传多样性分析、聚类分析和主成分分析。结果表明,9对SSR引物共检测到108个等位基因,等位基因数(Na)、有效等位基因数(Ne)、观测杂合度(Ho)、期望杂合度(He)、多态信息含量(PIC)、Shannon’s信息指数分别为12个、4.620个、0.678、0.651、0.629、1.568;聚类结果显示121份野生枸杞种质可划分为5个亚群,群体遗传结构表明当K=5时,121个种质划分为5个亚群,来自宁夏的NXGY-02、NXGY-032个种质与来自内蒙古的6个种质之间的遗传距离最远,亲缘关系较远,来自青海省、新疆、甘肃省及内蒙古的其余野生枸杞种质之间存在广泛的基因交流现象;宁夏、内蒙古的野生枸杞种质具有独立起源进化的可能,而新疆、甘肃省、青海省的种质与宁夏种质亲缘关系较近,很可能属于同一起源。

改进QuEChERS-UPLC-MS/MS法测定新疆枸杞中31种农药残留

目的:建立QuEChERS-UPLC-MS/MS法测定新疆枸杞中31种农药残留的方法。方法:样品经改进后的QuEChERS法净化后,经正负离子分段扫描和多反应监测模式检测,基质匹配标准溶液外标法定量。结果:在0.001~0.200 mg·L^(-1),31种目标化合物的线性相关系数均>0.996。回收率为70.2%~111.4%,相对标准偏差≤11.3%,检出限为0.01~7.97μg·kg^(-1)。结论:该方法快速、简便、准确,具有较强的适用性,可用于含糖量高的枸杞试样中多种农药残留的快速筛查与确证。

宁夏枸杞产业质量标准发展的现状、问题及对策

通过对比分析现行标准数量、结构布局、标准时效性、适应性、产品标准等阐述枸杞标准现状,分析宁夏枸杞产业在标准化及品牌化方面存在的不足,针对存在问题提出促进宁夏枸杞产业标准发展的5条建议,以期为我国枸杞产业总体质量标准水平提升提供参考。

《诗经》“杞”释说——兼辨《名医别录》之“榉”

《诗经》与植物“杞”相关的载记共有7处,同名异物的存在使得其整体类分与具体所指具有复杂性,诸家对此亦颇具分歧。综合观之,《诗经》之“杞”可分为3类:第一类是《四牡》《湛露》《四月》之“杞”,其为枸杞(Lycium chinense Miller);第二类是《将仲子》之“杞”,其指旱柳(Salix matsudana Koidz.);第三类是《南山有台》《杕杜》《北山》之“杞”,其是枫杨(Pterocarya stenoptera C.DC.)。《名医别录》之“榉”即为枫杨,《中国植物志》误将之释作榉树[Zelkova serrata(Thunb.)Makino]。

基于HPLC指纹图谱及聚类分析对不同产地枸杞质量评价研究

基于指纹图谱与系统聚类分析方法对不同产地枸杞进行质量评价。利用高效液相色谱法(HPLC)建立8个不同产地枸杞的指纹图谱,使用相似度评价软件及SPSS19.0软件进行数据处理,对不同产地的枸杞样品进行分析。结果表明,不同产地枸杞的黄酮类物质指纹图谱具有专属性,其中中宁、青海、新疆3个产地的枸杞指纹图谱相似度较高,分别为1、0.963、0.956,聚类分析也将该三个产地的枸杞聚在一起。青海、新疆产枸杞与中宁枸杞质量相似,可以作为枸杞药材新来源。HPLC指纹图谱结合聚类分析法能够对枸杞的产地识别及质量评价提供参考。

宁夏枸杞体外抗氧化机理研究

目的:研究宁夏枸杞中主要的4种抗氧化物质(枸杞多糖、黄酮、类胡萝卜素和VC)的抗氧化能力,分析它们对枸杞果实总抗氧化能力的贡献率,探索宁夏枸杞果实的体外抗氧化机理。方法:采用FRAP法、DPPH法、ABTS法和清除羟自由基法4种体外抗氧化方法来评价枸杞鲜果的总抗氧化能力。结果:4种方法测得枸杞黄酮对枸杞果实总抗氧化能力的贡献率均大于96%,VC对枸杞果实总抗氧化能力的贡献率均小于4%,高质量浓度的枸杞多糖(642.7170 mg/L)和类胡萝卜素(109.9280 mg/L)检测不到抗氧化能力。结论:枸杞黄酮的抗氧化能力决定着枸杞果实的体外抗氧化能力。

宁夏枸杞的道地性研究

宁夏是宁夏枸杞唯一的道地性产地。本文从地学的角度 ,通过数理统计的方法 ,对宁夏枸杞内在品质、影响宁夏枸杞的地球化学元素、影响宁夏枸杞品质的土壤地质背景条件等 3个方面综合论述了其道地性的原因 。

枸杞多糖提取条件优化及体外抗氧化活性研究

为了优化枸杞(Lycium chinense)中多糖的提取工艺并研究其抗氧化活性,以宁夏枸杞为原料,以水提醇沉为提取方法来研究其提取工艺。通过单因素试验,考察不同料液比、浸提温度、浸提时间、浸提p H、提取次数、醇沉时间、醇沉时乙醇的加入量等诸多因素来优化枸杞多糖的提取率;通过抗脂质过氧化能力的测定以及对超氧阴离子自由基清除作用的测定对枸杞多糖的抗氧化能力进行研究。结果表明,在提取枸杞多糖时,料液比为1∶35(g∶m L),温度为90℃,p H为11,浸提3 h,提取2次,合并提取液,加3倍体积95%乙醇沉淀6 h时,提取效果最佳,提取率为18.56%。另外,在提取时,采用超声波辅助提取30 min,可提高多糖提取率。抗氧化结果显示,枸杞多糖具有一定的抗氧化活性。

响应面法优化枸杞叶粗多糖提取纯化工艺及其降血糖活性

采用响应面法对枸杞叶粗多糖提取中的料液比、提取温度和提取时间进行优化,确定各因素最佳的水平组合。结果显示,在料液比1:47.2,提取温度72.9℃、提取时间2.2h条件下,枸杞叶粗多糖提取物中多糖的含量最高,为(7.24±0.41)%。采用D101大孔树脂和DEAE-52阴离子交换柱对枸杞叶粗多糖进行纯化,纯化后的精多糖中多糖含量为92.5%。用α-D-吡喃葡萄糖苷(p-NPG)法对枸杞叶精多糖的α-糖苷酶抑制活性进行了测定,1mg/mL枸杞叶精多糖对5U/mL的糖苷酶抑制率达到93.7%,而阳性对照药阿卡波糖抑制率仅为21.7%。

人参、黄芪、枸杞子对老年大鼠心肌β受体的影响

用放射配基结合分析法测定心肌β受体，观察老年大鼠心肌β受体的变化及中药人参、黄芪、枸杞子的影响。结果表明，心肌β受体最大结合容量（Bmax）在3、8、15月龄组间无显著差异，26月龄组显著降低。人参、黄芪、枸杞子对15月龄大鼠心肌β受体Bmax无明显影响，人参和枸杞子可显著提高26月龄大鼠心肌β受体Bmax，黄芪虽可使26月龄大鼠心肌β受体Bmax有所升高，但无显著性差异。表明衰老大鼠心肌β受体密度降低，中药可使之显著升高。提示对心肌β受体数目的调节是这些中药发挥抗衰老作用的分子基础之一。

地骨皮化学成分的研究

从山西产地骨皮（枸杞Ｌｙｃｉｕｍｃｈｉｎｅｎｓｅ根皮）中分离鉴定了４个化合物，为β－谷甾醇、盐酸甜菜碱、莨菪亭和阿魏酸二十八酯。

青海产三种特色果实多糖及其活性比较

用水提醇沉法提取了青海产宁夏枸杞、黑枸杞、西伯利亚白刺三种特色果实的水溶性总多糖,利用理化方法测定并比较了三种多糖的理化性质及其抗氧化活性和对腹腔巨噬细胞NO分泌功能的影响。三种植物多糖分子量范围为130000~2350000;单糖组成都含有甘露糖、鼠李糖、葡萄糖醛酸、半乳糖醛酸、葡萄糖、半乳糖、木糖及阿拉伯糖,但半乳糖醛酸、葡萄糖和阿拉伯糖的相对含量差异明显;三者的糖醛酸含量及蛋白质含量也有明显差异。活性测定结果表明,西伯利亚白刺多糖具有明显的DPPH清除能力,其IC50值达到0.19 mg/m L,表现出较强的抗氧化活性,而黑枸杞与宁夏枸杞多糖的活性较弱;三种多糖均可显著增强小鼠腹腔巨噬细胞的NO分泌水平,显示一定的免疫增强活性。