Based on non-targeted metabolomics for differential components screening of Rosae Chinensis Flos and Rosae Rugosae Flos and their quality evaluation

Background:Rosa chinensis Jacq.and Rosa rugosa Thunb.are not only of ornamental value,but also edible flowers and the flower buds have been listed in the Chinese Pharmacopoeia as traditional medicines.The two plants have some differences in efficacy,but the flower buds are easily confused for similar traits.In addition,large-scale cultivation of ornamental rose flowers may lead to a decrease in the effective components of medicinal roses.Therefore,it is necessary to study the chemical composition and make quality evaluation of Rosae Chinensis Flos(Yueji)and Rosae Rugosae Flos(Meigui).Methods:In this study,40 batches of samples including Meigui and Yueji from different regions in China were collected to establish high-performance liquid chromatography fingerprints.Then,the fingerprints data was analyzed using principal component analysis,hierarchical cluster analysis,and partial least squares discriminant analysis analysis chemometrics to obtain information on intergroup differences,and non-targeted metabolomic techniques were applied to identify and compare chemical compositions of samples which were chosen from groups with large differences.Differential compounds were screened by orthogonal partial least-squares discriminant analysis and S-plot,and finally multi-component quantification was performed to comprehensively evaluate the quality of Yueji and Meigui.Results:The similarity between the fingerprints of 40 batches roses and the reference print R was 0.73 to 0.93,indicating that there were similarities and differences between the samples.Through principal component analysis and hierarchical cluster analysis of fingerprints data,the samples from different origins and varieties were intuitively divided into four groups.Partial least-squares discriminant analysis analysis showed that Meigui and Yueji cluster into two categories and the model was reliable.A total of 89 compounds were identified by high resolution mass spectrometry,mainly were flavonoids and flavonoid glycosides,as well as phenolic acids.Eight differential components were screened out by orthogonal partial least-squares discriminant analysis and S-plot analysis.Quantitative analyses of the eight compounds,including gallic acid,ellagic acid,hyperoside,isoquercitrin,etc.,showed that Yueji was generally richer in phenolic acids and flavonoids than Meigui,and the quality of Yueji from Shandong and Hebei was better.It is worth noting that Xinjiang rose is rich in various components,which is worth focusing on more in-depth research.Conclusion:In this study,the fingerprints of Meigui and Yueji were established.The chemical components information of roses was further improved based on non-targeted metabolomics and mass spectrometry technology.At the same time,eight differential components of Meigui and Yueji were screened out and quantitatively analyzed.The research results provided a scientific basis for the quality control and rational development and utilization of Rosae Chinensis Flos and Rosae Rugosae Flos,and also laid a foundation for the study of their pharmacodynamic material basis.

Identification of key genes regulating the synthesis of quercetin derivatives in Rosa roxburghii through integrated transcriptomics and metabolomics

Rosa roxburghii fruit is rich in flavonoids, but little is known about their biosynthetic pathways. In this study, we employed transcriptomics and metabolomics to study changes related to the flavonoids at five different stages of R. roxburghii fruit development. Flavonoids and the genes related to their biosynthesis were found to undergo significant changes in abundance across different developmental stages, and numerous quercetin derivatives were identified. We found three gene expression modules that were significantly associated with the abundances of the different flavonoids in R. roxburghii and identified three structural UDP-glycosyltransferase genes directly involved in the synthesis of quercetin derivatives within these modules. In addition, we found that RrBEH4, RrLBD1 and RrPIF8could significantly increase the expression of downstream quercetin derivative biosynthesis genes. Taken together,these results provide new insights into the metabolism of flavonoids and the accumulation of quercetin derivatives in R. roxburghii.

Effect of fermented Rosa roxburghii Tratt fruit juice on gut microbiota in a dextran sulfate sodium-induced colitis mouse model

This study mainly investigated the regulatory effect of Rosa roxburghii Tratt fruit juice fermented by Lacticaseibacillus paracasei SR10-1(LAB-RRTJ)on modulating gut microbiota in dextran sulfate sodium(DSS)-induced ulcerative colitis in mice.Compared to control group,DSS induction decreased body weight of mice,indexes of Shannon,Simpson,Chao1 and Faith_pd,and increased disease activity index(DAI)and levels of interleukin 1β(IL-1β),IL-6,tumor necrosis factorα(TNF-α)and interferon-γ(IFN-γ);And this induction also led to an increase in Proteobacteria,Verrucomicrobia and Actinobacteria at phylum level,harmful bacterial species richness at genus level,and relative richness of S.sciuri,Desulfovibrio C21_c20,R.gnavus and Akkermansia muciniphila at species level,and a decrease in Firmicutes at phylum level and relative richness of B.acidifaciens in mice.LAB-RRTJ increased body weight of mice with DSS induced ulcerative colitis(UC)and indexes of Shannon,Simpson,Chao1 and Faith_pd,reduced DAI and the content of four infl ammatory factors and improved gut microbiota imbalance in DSS induced UC mice.Besides,the number of operational taxonomic units(OTUs)increased,α-diversity andβ-diversity were restored and similar to those in mice in the control group after LAB-RRTJ treatment.Compared with the positive drug treatment group,LAB-RRTJ has a better effect on regulating gut microbiota diversity in colitis mice.Correlation analysis showed that infl ammatory factors were positively correlated with harmful bacteria and negatively correlated with beneficial bacteria which commonly found in some colitis mice.Taken together,our study demonstrated that LAB-RRTJ could alleviate DSS-induced colitis in mice through the modulation of infl ammatory cytokines and gut microbiota composition.

Mechanism of Rosae Rugosae Flos flavonoids in the treatment of hyperlipidemia and optimization of extraction process based on network pharmacology

This study aims to identify a natural plant chemical with hypolipidemic effects that can be used to treat high cholesterol without adverse reactions.Through network pharmacology screening,it was found that Rosae Rugosae Flos(RF)flavonoids had potential therapeutic effects on hyperlipidemia and its mechanism of action was discussed.TCMSP and GeneCards databases were used to obtain active ingredients and disease targets.Venn diagrams were drawn to illustrate the findings.The interaction network diagram was created using Cytoscape 3.8.0 software.The PPI protein network was constructed using String.GO and KEGG enrichment analysis was performed using Metascape.The results revealed 2 active flavonoid ingredients and 60 potential targets in RF.The key targets,including CCL2,PPARG,and PPARA,were found to play a role in multiple pathways such as the AGE-RAGE signaling pathway,lipid and atherosclerosis,and cancer pathway in diabetic complications.The solvent extraction method was optimized for efficient flavonoid extraction based on network pharmacology prediction results.This was achieved through a single factor and orthogonal test,resulting in an optimum process with a reflux time of 1.5 h,a solid-liquid ratio of 1:13 g/mL,and an ethanol concentration of 50%.

Exploring the mechanism of Crocus sativus and Rosa rugosa for the treatment of coronary heart disease based on network pharmacology and molecular docking

Coronary atherosclerotic heart disease(CHD)is the main type of cardiovascular disease.The efficacy of Uyghur drug compound Saffron formula in CHD has been clinically proven.However,the underlying mechanism remains unclear.In this study,researchers investigated the active ingredients and mechanism of action of Crocus sativus and Rosa rugosa in the treatment of CHD by network pharmacology and molecular docking techniques,collected target information with the help of TCMSP,GEO,GeneCards,and other databases,constructed protein-protein interaction(PPI)network diagrams by STRING database,performed GO and KEGG pathway enrichment analysis on common targets,and finally molecularly docked the active ingredients with core targets.C.sativus-R.rugosa have a variety of polyphenol compounds,a total of 12 active ingredients,including quercetin and kaempferol,were screened.The first three targets intersected with the core targets of CHD as AKT1,TNF,and IL-1B.Enrichment results of KEGG pathway showed that C.sativus-R.rugosa against CHD involved atherosclerosis pathways.The molecular docking results showed that quercetin and kaempferol were well bound to the core targets,and it was speculated that these components might be the main active ingredients for the treatment of CHD.The potential mechanism of action of C.sativus-R.rugosa for the treatment of coronary heart disease was initially revealed.

Structural characteristics,anticoagulant and antithrombotic mechanism of a novel polysaccharide from Rosa Chinensis Flos

This is the first report on a polysaccharide(RCJ2-Ib)isolated from Rosa Chinensis Flos.RCJ2-Ib was obtained through the extraction with water,precipitation with ethanol,separation with DEAE-52 column and purification with DEAE-Sepharose Fast Flow column and Sephadex G100 column.GC,FT-IR and NMR analyses revealed that RCJ2-Ib(3.3 k Da)was a 1,4-linked polymannuronic acid containing substantialβ-Danomers units.The anticoagulant effect of RCJ2-Ib evaluated by using rabbit ear venous blood and an acute blood stasis rat model showed that RCJ2-Ib had obvious anticoagulant activity in regulating endogenous and exogenous coagulation pathways and reducing serum thromboxane B2 and endothelin-1.In addition,RCJ2-Ib could also increase the number of Lactobacillus and Escherichia coli.As a result,RCJ2-Ib has the potential to inhibit thrombosis and maintain the intestinal environment.

Polysaccharide extract from Rosa laevigata fruit attenuates inflammatory obesity by targeting redox balance and gut interface in high-fat diet-fed rats

Low-molecular-weight polysaccharides(RLPs)extracted from Rosa laevigata fruits showed multiple biofunctions in Asia.This study aimed to investigate its anti-inflammatory obesity effect in high-fat dietfed rats and further elucidated the underlying molecular mechanism by multi-omics methods.The results showed that RLPs administration had significantly restored immune organ indexes and reduced body weight gain.RNA-seq revealed that the effect of RLPs was partially attributed to its regulation on PPARs signaling by increasing the expressions of Scd,Acox3 and Hmgcs2,and on other redox-related pathways by decreasing the expressions of Cyp2 e1,Il1-r1 and Lbp.Moreover,16 S rRNA sequencing coupled with metagenome sequencing showed that RLPs had significantly reduced the ratio of Firmicutes/Bacteroidetes from 8.01 to2.37,and significantly increased the relative abundances of Alistipes,Prevotella,and Akkermansia from 0.36%,1.10%and 2.61%to 0.65%,2.37%and 4.42%,respectively.Spearman correlation analysis result indicated that the abundances of Lachnospiraceae,Prevotella and Bacteroidales were significantly negatively correlated with obesity phenotype,liver function and inflammatory factors.These results revealed that RLPs exerted significant anti-inflammatory obesity property partially via regulation on gut microbiota interface and the redox balance.Therefore,RLPs could be a promising functional food resource with the potential for redox imb alance-related diseases chemoprevention.

Exploring the Protective Effect of the Ethanolic Extract of Rosa laevigata Michx.Fruit on Rats with Mesangial Proliferative Glomerulonephritis Based on NLRP3 Inflammasome Pathway

Objective:To investigate the effect of the ethanolic extract of Rosa laevigata Michx.fruit on rats with mesangial proliferative glomerulonephritis based on the NLRP3 inflammasome pathway.Methods:Thirty Wistar rats were divided into three groups,a blank control group,a diabetic nephropathy(DN)model group,and an ethanolic extract intervention group,according to the random number table method,with 10 rats in each group.One day before the experiment,basic feeding was initiated for all the rats;the changes in activity and weight of each group of rats were observed and recorded after 7 d,and a rat model of renal function injury was established after 1 d.Results:Compared with the control group,the model group had significantly higher kidney/body ratio,24 h urine protein,serum creatinine(SCr),blood urea nitrogen(BUN),glomerular mesangial cell(GMC)count,and extracellular matrix(ECM)positive area ratio(P<0.05);the same indicators were significantly lower in the intervention group than in the model group(P<0.05).The NLRP3 inflammasome pathway in renal intrinsic cells was activated in the intervention group.The overactivation of NLRP3 inflammasome is known to promote interleukin(IL)-1βrelease,which was inhibited in the intervention group.Conclusion:The ethanolic extract of Rosa laevigata Michx.fruit has a protective effect on renal intrinsic cells and may be related to NLRP3 inflammasome pathway,suggesting that the fruit of Rosa laevigata Michx.has a potential role in protecting renal intrinsic cells from inflammatory damage.NLRP3 inflammasomes are involved in the development of various chronic inflammatory diseases,such as acute and chronic glomerulonephritis and renal fibrosis.

Isolation and Screening of Black Spot-Resistant Endophytic Fungi Strains from Rosa chinensis

[Objective] This study aimed to isolate and screen black-spot-resistant en- dophytic fungi strains. [Method] Various species of healthy Rosa chinensis were col- lected from Xi＇an, Xianyang, Baoji and Weinan City in Shaanxi Province. Endophytic fungi was isolated from stems and leaves, and purified by 2-3 times of inoculation to screen endophytic fungi antagonistic to Marssonina rosae with modified punching method. [Result] Samples collected from Xianyang exhibited the highest colonization rate and isolation rate; endophytic fungi strains isolated from stems presented the highest colonization rate and isolation rate compared with leaves. A total of 67 en- dophytic fungi strains were isolated from Rosa chinensis, including 3 black spot-re- sistant strains which were all derived from Baoji. [Conclusion] This study laid the foundation for further screening candidate strains of biocontrol fungi and environ- ment-friendly fungal biological control.

岷江干旱河谷黄蔷薇(Rosa hugonis)生长与繁殖特征及其空间差异

对乡土植物生长繁殖特征的认识是制定合理的植被恢复及管理措施的基础和依据。通过大面积的野外踏查和样方调查,分析了岷江干旱河谷乡土灌木-黄蔷薇(Rosa hugonis)的分布、生长繁殖特征及空间差异。发现:(1)黄蔷薇主要分布于岷江干旱河谷的茂县段,其植株株高、基径、年龄、丛冠幅、丛的萌生数、丛干重、丛生物量和密度等生长特征存在明显空间差异,除丛萌生数和密度之外海拔对其他各参数都有较大的影响,这些参数的值在高海拔地段大于低海拔地段;(2)黄蔷薇在干旱河谷丛平均萌生数为6.48±0.97株/丛,丛的萌生能力较强,且其受砍伐后具有很强的再生能力;萌生株年龄结构大致为金字塔型,最大植株年龄为25 a;(3)干旱河谷黄蔷薇的结实量较大,平均值为(404.96±106.91)果/丛,结实特征表现出明显的空间差异性;(4)各生长与结实参数关系密切,影响结实量的最主要的生长因素是冠幅和丛干重。研究结果表明:黄蔷薇适应干旱河谷的环境条件,具有较广泛的分布,生长和和繁殖状况良好,是可供选择的干旱河谷恢复物种。

Chemical Constituents from Fruits of Rosa davidii

From the fruit of Rosa davidii Crep., eleven compounds were isolated and identified by spectral evidence, viz. 2 alpha, 3 beta, 19 beta -trihydroxyl-olean-12-en-28-oic acid (1), 2 alpha, 3 beta -dihydroxyl-urs-28 (13)-lactone (2), arjunic acid (3), euscaphic acid (4), 2 alpha, 3 beta -dihydroxyl-urs-12-en-28-oic acid (5), oleanolic acid (6), kaempferol (7), tiliroside (8), quercetin (9), daucosterol (10) and beta -sitosterol (11). Among them, 1 and 2 were new compounds.

ROSA法快速检测鲜奶中的四环素类和β-内酰胺类药物残留

本文主要探讨应用ROSA法同时快速检测新鲜牛奶中的四环素类和β-内酰胺类药物残留。评价ROSA法的灵敏度和特异性,同时验证ROSA法的检测稳定性,该法10min可出检测结果。

ROSA法快速检测UHT奶中的四环素类和β-内酰胺类药物残留

主要探讨应用ROSA法同时快速检测UHT奶中的四环素类和β-内酰胺类药物残留。评价了ROSA法的灵敏度和特异性,验证了ROSA法的检测稳定性,10 min可出检测结果。

ZFN,TALEN和CRISPR/Cas9在小鼠Rosa26基因定点整合外源基因的效率比较

哺乳动物黑色素的合成依赖于酪氨酸的氧化作用,而酪氨酸酶(Tyr)是催化酪氨酸氧化反应的关键酶,当外源Tyr基因整合进白毛色小鼠基因组中,会使它获得黑色素合成的能力,表现出与原来不同的毛色表型。为方便、快捷地获得Tyr基因整合的小鼠,构建了一个无启动子的pTyr-2A-DsRed同源重组质粒供体,选择Rosa26的第一个内含子作为外源基因整合的靶位点,设计了切割位点几乎一致的ZFN、TALEN和CRISPR/Cas9系统。通过流式对比分析C 2C 12细胞中红色荧光蛋白DsRed的表达水平,比较了3种基因组编辑工具介导的外源基因定点整合效率,结果发现CRISPR/Cas9的效率最高,在此基础上,利用CRISPR/Cas9将供体整合到小鼠胚胎干细胞中,筛选单细胞克隆进行囊胚腔注射和胚胎移植,获得一只存活的嵌合体小鼠,表现出白毛中夹杂黑毛的表型,表明整合到小鼠Rosa26的Tyr基因可以正常表达。

ROSA机器人辅助下立体定向活检术在脑内病变诊断中的作用

目的探讨ROSA机器人引导下立体定向活检术方法及其在脑内病变诊断中的作用。方法回顾性分析2015年6月至2016年9月收治的18例脑内病变患者的临床资料,所有患者均在ROSA机器人辅助下行立体定向活检术。结果 18例活检术阳性率为100%,其中淋巴瘤7例,低级别胶质瘤1例,胶质母细胞瘤4例,间变星形细胞瘤2例,弥漫性星形细胞瘤1例,病毒性脑炎1例,炎性脱髓鞘1例,亚急性出血性脑梗死1例。18例平均手术时间为80 min。术后均未发生永久性神经功能障碍,也无颅内感染;1例术后复查CT示无症状性颅内出血。结论 ROSA机器人辅助下立体定向活检术具有安全、精确、省时等优势,可为脑内病变诊断提供有力技术支持。

A Genetic Transformation System for Rosa multiflora Thunb. var. cathayensis Rehd. et Wils through Callus Induction

An efficient genetic transformation system is a preparation for Rosa multi-flora Thunb. var. cathayensis Rehd. et Wils to diversify its flower color through ge-netic engineering. We firstly optimized the explants and culture conditions on callus induction, hormone concentrations and dark period of culture time on bud differentia-tions in particular, with sterilized seedlings to establish the regeneration system of R. multiflora. It showed that callus induction frequency reached 100% after the ex-plants being cultured in dark for 21 d when MS was chosen to be the initial culture medium. The bud differentiation rate was 48% after cal i being cultured under dark for 8 d on MS medium supplemented with TDZ （1.5 mg/L） and NAA （0.05 mg/L）. The cal i was used as the explants that were infected with Agrobacterium tumefa-ciens harboring a DFR-RNAi construct. The transformation rate reached as high as 50%. The establishment of a highly efficient rose gene transformation system out-lined in this report is prerequisite for genetic improvement in rose flower colors.

Development of an in vitro culture system of Rosa spp.Plants

[Objective] This study was to develop an in vitro tissue culture system of Rosa spp.[Method] Using ten species of Rosa spp.plants as experimental materials,different combinations of hormones were designed to establish their in vitro tissue culture system with the stem segments as explants.[Result] All ten tested varieties germinated when the nodal segment explants were cultured on the sprouting medium MS＋ 0.5 mg/L BA ＋0.01 mg/L NAA and grew vigorous shoots,and the sprouting rate was up to 70%.Of the ten tested rose varieties,each has a respective optimal proliferation medium,and the multiplication rates for all the varieties reached 3.0%.The axillary buds were vigorous and normal in leaf color.The optimal medium for rooting and acclimation was 1/2MS medium containing 0.1 mg/L or 0.2 mg/L NAA,in which the rooting frequency reached 90%-100% and the root system was developed.After acclimation and transplant,the survival rate was as high as 95%.[Conclusion] An in vitro tissue culture system of Rosa spp.has been established in this study,which lays foundation for the molecular breeding of Rosa spp.

ROSA法快速检测婴儿奶粉中四环素类和β-内酰胺类药物残留

为了建立同时快速检测婴儿奶粉中四环素类和β-内酰胺类药物残留的方法,试验采用RO-SA法测定了市购婴儿奶粉中的药物残留。结果表明:ROSA法的灵敏度和特异性均能达到检测要求,15 min可获得检测结果。说明ROSA检测方法可以用来检测婴儿奶粉中的药物残留。

ROSA辅助治疗高血压脑出血的疗效研究

目的:探讨立体定向手术机器人(ROSA)辅助治疗高血压脑出血的疗效和安全性。方法:收治高血压脑出血患者158例,分为ROSA辅助治疗组(ROSA组)与框架辅助治疗组(框架组),比较两组治疗效果。结果:框架组手术时间、术后拔管时间和术后再出血率均显著高于ROSA组(P<0.05)。结论:ROSA辅助治疗高血压脑出血的疗效显著,安全性高。

耳颞神经阻滞(ANB)在ROSA机器人辅助下颅内电极植入术后镇痛中的应用

目的评估选择性耳颞神经阻滞(auriculotemporal nerve block,ANB)应用于ROSA机器人辅助下颅内电极植入术术后镇痛的效果。方法60例接受ROSA机器人辅助下颅内电极植入术的患者,年龄13~65岁,16<BMI≤29.9,ASAⅠ~Ⅱ级。采用随机数字表法分为耳颞神经阻滞组(ANB组,n=30)和静脉患者自控镇痛组(PCIA组,n=30)。ANB组在手术结束后用0.5%罗派卡因2~3 mL在超声引导下阻滞双侧耳颞神经,术后留置导管,每8 h通过留置导管推注0.5%罗哌卡因2 mL以维持神经阻滞效果;PCIA组术后配制镇痛泵,2μg/kg舒芬太尼+2μg/kg右美托咪啶+昂丹司琼8 mg+0.9%氯化钠注射液,稀释至100 mL。术后4、12、24和48 h记录患者的简化McGill疼痛评分和不良反应,包括恶心呕吐、皮肤瘙痒、呼吸抑制、低血压、心动过缓、局麻药中毒等。结果与PCIA组比较,ANB组术后4、12、24和48 h的McGill疼痛评分差异无统计学意义,ANB组的不良反应与12 h内张口咬合疼痛发生率明显低于PCIA组。结论超声引导下选择性ANB是ROSA机器人辅助下颅内电极植入术术后镇痛的有效方法,较PCIA镇痛更具优势。