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.

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%.

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.

Exogenous calcium enhances the physiological status and photosynthetic capacity of rose under drought stress

Drought(water shortage)can substantially limit the yield and economic value of rose plants(Rosa spp.).Here,we characterized the effect of exogenous calcium(Ca^(2+))on the antioxidant system and photosynthesis-related properties of rose under polyethylene glycol 6000(PEG6000)-induced drought stress.Chlorophyll levels,as well as leaf and root biomass,were significantly reduced by drought;drought also had a major effect on the enzymatic antioxidant system and increased concentrations of reactive oxygen species.Application of exogenous Ca^(2+)increased the net photosynthetic rate and stomatal conductance of leaves,enhanced water-use efficiency,and increased the length and width of stomata following exposure to drought.Organ-specific physiological responses were observed under different concentrations of Ca^(2+).Application of 5 mmol·L^(-1)Ca^(2+)promoted photosynthesis and antioxidant activity in the leaves,and application of 10 mmol·L^(-1)Ca^(2+)promoted antioxidant activity in the roots.Application of exogenous Ca^(2+)greatly enhanced the phenotype and photosynthetic capacity of potted rose plants following exposure to drought stress.Overall,our findings indicate that the application of exogenous Ca^(2+)enhances the drought resistance of roses by promoting physiological adaptation and that it could be used to aid the cultivation of rose plants.

The MADS-box gene RhAGL6 plays a master role in regulating the receptacle malformation in rose at low temperature

Low temperature usually results in the developmental deformity of flower organs,immensely affecting the quality of rose flowers.However,it's largely unknown about the regulatory mechanisms activated by low temperature.Here,we used a low temperature-sensitive Rosa hybrida cv.‘Peach Avalanche’to screen a MADS-box gene RhAGL6 via conjoint analysis between RNA sequencing(RNA-seq)and whole-genome bisulfite sequencing(WGBS).Furthermore,we found that low temperature induced the hypermethylation and elevated histone 3 lys-27 trimethylation(H3K27me3)level on the RhAGL6 promoter,leading to decreased RhAGL6 expression.In addition,RhAGL6 silencing resulted in the formation of abnormal receptacles.We also found that the levels of gibberellins(GA3)and abscisic acid(ABA)in the receptacle under low temperature were lower and higher,respectively,than under normal temperature.Promoter activity analysis revealed that GA3 significantly activated RhAGL6 promoter activity,whereas ABA inhibited it.Thus,we propose that RhAGL6 regulates rose receptacle development by integrating epigenetic regulation and phytohormones signaling at low temperature.

Vav^(Cre)介导的荧光报告系统对小鼠小胶质细胞的标记效率

目的:探讨Vav^(Cre)介导的荧光报告系统对小胶质细胞的标记效率。方法:将Vav^(Cre)/Rosa26REYFP fl/fl雄鼠与C57BL/6雌鼠杂交获得Vav^(Cre)/Rosa26REYFP^(fl/-)和Rosa26REYFP^(fl/-)子代。以发现雌鼠阴道栓的时间标记为E0.5,流式细胞术检测EYFP在E7.5~E8.25、E10~E10.5、E12~E12.5、E14~E14.5小鼠胚胎和新生小鼠小胶质细胞中的表达。结果:Vav^(Cre)/Rosa26REYFP^(fl/-)组E10~E10.5、E12~E12.5、E14~E14.5胚胎和新生小鼠小胶质细胞中EYFP阳性率高于Rosa26REYFP^(fl/-)组(P<0.001),且均>95.000%。Vav^(Cre)/Rosa26REYFP^(fl/-)E7.5~E8.25小鼠胚胎中EYFP阳性率为(0.059±0.009)%。结论:以Vav^(Cre)介导的荧光报告系统可以稳定、高效地标记小鼠小胶质细胞,可以用于动态监测小胶质细胞的发育。

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.

Network pharmacology research and experimental verification of Huangqi(Astragalus Radix)and Jinyingzi(Rosae Laevigatae Fructus)in treating benign prostatic hyperplasia

Objective This study aimed to analyze the mechanism of action of Huangqi(Astragalus Radix,HQ)-Jinyingzi(Rosae Laevigatae Fructus,JYZ)in the treatment of benign prostatic hyperplasia(BPH)based on network pharmacology and to verify the prediction through animal experimentation.Methods Based on the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine(BATMAN-TCM)databases,and literature,the active components and related target genes of HQ and JYZ were screened.The BPH target genes were screened based on the DisGeNET and GeneGards databases,and Excel was used to merge and remove duplicates.The Perl language was used to obtain drug-BPH target genes by intersecting shared target genes.A drug-component-target gene network diagram was constructed using Cytoscape software.The drug-BPH intersection target genes were inputted into the STRING database,and the key target genes were selected according to the degree algorithm.The output formed the basis for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses to determine the potential mechanism of HQ and JYZ in BPH treatment.High,medium,and low doses of HQ and JYZ extract were used to intervene in BPH rats,and then the prostate volume,wet weight,and prostate index of the BPH rats were determined.Changes in prostate histopathology and microvessel density(MVD)were evaluated using immunohistochemistry,and the optimal HQ and JYZ extract dose was confirmed.Finally,the optimal dose was used to intervene in a BPH rat model,and AKT1 and VEGF expressions were examined by immunohistochemistry.Results Based on network pharmacology,33 active components and 772 target genes were identified from HQ and JYZ,along with 817 BPH target genes and 112 drug-BPH common target genes.Among them were 10 key target genes,including AKT1,JUN,MAPK1,IL-6,TNF,ESR1,and VEGFA.KEGG enrichment analysis revealed 135 signaling pathways,including PI3K/AKT,IL-17,TNF,p53,MAPK,VEGF,JAK-STAT,and NF-κB pathways.The animal experiment showed that HQ and JYZ significantly improved prostate volume,wet weight,prostate index,and prostate histopathology of BPH rats,reducing MVD.In addition,HQ and JYZ inhibited the expression of AKT1 and VEGF in the prostate tissue of rats,promoted epithelial cell apoptosis,and inhibited angiogenesis,consistent with the prediction.Conclusion The combination of HQ and JYZ is effective for BPH therapy through multi-compound and multi-target collaboration.Its possible mechanism in treating BPH includes regulation of AKT1,VEGF protein,PI3K/Akt,and VEGF signaling pathways related to apoptosis,angiogenesis,and inflammation,with potential for clinical use and research.

In vivoand in vitroanti-arthritic efficacy of a herbal formula consisting of Rosae Multiflorae Fructus and Lonicerae Japonicae Flos

OBJECTIVE Rheumatoid arthritis(RA)is the most common inflammatory autoimmune disease,affecting around 1% of the world population.Toll-like receptor 4(TLR4)signalling has been found to be involved in the pathogenesis of RA.It is a potential therapeutic target for RA treatment.A herbal formula(RL)consisting of Rosae Multiflorae Fructus and Lonicerae Japonicae Flos has traditionally been used in treating various inflammatory disorders.In this study,we would evaluate the anti-arthritic effect of RL on collagen-induced arthritis(CIA)in rats and investigate the involvement of TLR4 signaling in the mode of action of RL in vivo and in vitro.METHODS In vivo anti-arthritic efficacy was evaluated using CIA rats induced by bovine typeⅡ collagen.The treatment groups were treated with various concentrations of RL or positive control indomethacin for 35 d.Clinical signs(hind paw volume and arthritis severity scores),changes in serum inflammatory mediators,histological and radiographic changes of joints were investigated.Spleens and peritoneal macrophages were used to determine the effects of RL on innate and adaptive immune responses in CIA rats.The involvement of TLR4 signalling pathways in the anti-arthritic effect of RL was examined in cartilage tissue of CIA rats,murine RAW264.7macrophages and human THP-1 monocytic cells.RESULTS The severity of arthritis in the CIA rats was significantly attenuated by RL.Histological score and radiographic score were efficiently improved by RL.RL could also dose-dependently inhibit pro-inflammatory cytokines in serum of CIA rats.RL significantly inhibited the production of various pro-inflammatory mediators,the expression and/or activity of the components of TLR4 signalling pathways in animal tissue and cell lines.CONCLUSION RL possesses anti-arthritic effect on collagen-induced arthritis in rats.The therapeutic effect of RL may be related to its inhibition on pro-inflammatory cytokines in serum.The inhibition of the TLR4/TAK1/NF-κB and TLR4/TAK1/MAPK pathways participate in the anti-arthritic effects of RL.This provides a pharmacological justification for the use of RL in the control of various arthritic diseases.Further investigation should be done to develop RL into a modern anti-arthritic agent.

Occurrence Factors and Comprehensive Prevention and Control of Chinese Rose Black Spot(Actinonema rosae)

In addition to conventional techniques,the prevention and control of Chinese rose black spot(Actinonema rosae)should be strengthened in the management of Chinese rose.The occurrence of Chinese rose black spot is closely related to plant residues,rainfall,cultivation conditions and varieties.On the basis of understanding the symptoms and occurrence regularity of Chinese rose black spot,combination measures of agricultural and chemical prevention and control should be taken.

The Study on the Action Mechanism of the Jinyingzi(Rosae Laevigatae Fructus)–Qianshi(Euryales Semen)Couplet Herbs on Membranous Nephropathy Based on Network Pharmacology

Objective Our objective was to explore the action mechanism of the Jinyingzi(Rosae Laevigatae Fructus)–Qianshi(Euryales Semen)couplet herbs in the treatment of membranous nephropathy(MN)based on network pharmacology.Methods The active ingredients and targets of Jinyingzi(Rosae Laevigatae Fructus)and Qianshi(Euryales Semen)were screened by systematic pharmacology database and analysis platform.Online Human Mendelian Genetic database and GeneCards database were used to retrieve MN-related targets.The active ingredient-related targets and MN disease targets were introduced into Venny 2.1,and Wayne diagram was drawn.The intersection targets were the potential targets of the Jinyingzi(Rosae Laevigatae Fructus)–Qianshi(Euryales Semen)couplet herbs in the treatment of MN.The protein interaction network of potential targets was constructed,and the core targets were screened with String platform.Metascape platform was used for functional enrichment analysis of gene ontology(GO)and pathway enrichment analysis of Kyoto Encyclopedia of Genes and Genomes(KEGG).The“herb-active ingredient-target-pathway”networks were drawn by using Cytoscape software,and the key components,targets,and signaling pathways were screened.Results A total of 8 active ingredients and 193 related targets in Jinyingzi(Rosae Laevigatae Fructus)and Qianshi(Euryales Semen)were screened out;a total of 1,621 targets of MN disease and 105 potential targets for the treatment of MN were obtained in the treatment with Jinyingzi(Rosae Laevigatae Fructus)–Qianshi(Euryales Semen)couplet herbs;40 core targets were screened by protein–protein interaction network topology analysis;a total of 1,978 results were obtained by GO function enrichment analysis,and 206 signal pathways were obtained by KEGG pathway enrichment analysis and screening.The network topology analysis of“herb-active ingredient-target-pathway”showed that the key components included quercetin,kaempferol,β-sitosterol,etc.;the key targets included protein kinase Bα(AKT),mitogen-activated protein kinase 1(MAPK1),B-cell lymphoma-2(BCL2),prostaglandin-endoperoxide synthase 2(PTGS2),etc.;the key pathways included advanced glycation end product/receptor of AGE signaling pathway,phosphatidyl inositol 3-kinase(PI3K)/AKT signaling pathway,MAPK signaling pathway,hypoxia-inducible factor-1 signaling pathway,Ras signaling pathway,nuclear factor kappa-B(NF-κB)signaling pathway,Toll-like receptors signaling pathway,p53 signaling pathway and vascular endothelial growth factor signaling pathway in the late stage of diabetic complications.Conclusion The Jinyingzi(Rosae Laevigatae Fructus)–Qianshi(Euryales Semen)couplet herbs can regulate PI3K/AKT,MAPK,NF-κB signaling pathways in MN by targeting proteins of AKT1,MAPK8,PTGS2 through key components of quercetin,β-sitosterol,and kaempferol,so as to inhibit the overexpression of inflammatory factors in renal tissues,regulate inflammatory response,and improve renal function.

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.

利用Cas9/sgRNA体系提高猪胎儿成纤维细胞基因组编辑效率

[目的]本文旨在利用Cas9/sgRNA体系,提高猪胎儿成纤维细胞的基因组编辑效率。[方法]针对猪ROSA26位点的启动子区(包含第一外显子区)设计2条sgRNA,将其构建在PX459-cas9-puro质粒上,以获得PX459-cas9-sgRNA质粒。将sgRNA靶序列连同原间隔序列(protospacer-adjacent motif,PAM)构建在RGS-CR质粒上,以获得RGS-sgRNA质粒。使用PX459-cas9-sgRNA质粒与RGS-sgRNA质粒系统共转染,筛选打靶效率较高的一条sgRNA,并使用该sgRNA对猪胎儿成纤维细胞(PEF)进行后续基因打靶试验。分别使用PX459-cas9-sgRNA质粒和Cas9/sgRNA 2种方法对PEF的靶位点进行基因敲除,将转染后的细胞提取基因组DNA,以桑格尔测序检测打靶效率。[结果]成功构建了打靶猪ROSA26靶序列的PX459-cas9-sgRNA质粒和含PAM序列的RGS-sgRNA,质粒测序结果显示序列插入正确;RGS-sgRNA筛选结果表明,sgRNA2的打靶效率极显著高于sgRNA1(P<0.01),且sgRNA2无脱靶效应,因此后续使用PX459-cas9-sgRNA2和Cas9/sgRNA2在PEF中进行基因打靶试验,并比较两者的基因打靶效率。在猪胎儿成纤维中,PX459-cas9-sgRNA2质粒的打靶效率为15.67%,Cas9/sgRNA2体系的打靶效率为33.33%。[结论]Cas9/sgRNA体系较PX459-cas9-sgRNA质粒打靶效率提高了约1倍,这对提高猪体细胞的基因编辑效率和促进基因编辑在猪上的应用有重要意义。

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.

应用CRISPR/Cas9技术构建Tet-on调控表达uPA的人源化肝细胞嵌合小鼠

目的构建Tet-on调控小鼠肝脏特异性表达uPA基因的NOD-SCID背景的转基因小鼠,并通过人肝癌细胞移植修复小鼠亚急性肝损伤进而构建人源化肝细胞嵌合小鼠。方法采用CRISPR/Cas9技术,在gRNA引导下,Cas9核酸内切酶于Rosa26基因位点切割并编辑,插入目的基因pTight-uPA-pA-Alb promoter-rtTA-pA表达框。使目的基因能跟随染色体稳定遗传,生物学功能保持稳定。新培育的转基因小鼠分为4组:诱导组(n=17)、移植组(n=16)、未诱导组(n=5)和空白对照组(n=5)。诱导组经强力霉素诱导3周,肝脏中表达的uPA能造成小鼠肝脏亚急性损伤;再通过人肝癌细胞移植修复。移植4周后,通过小鼠血清生化学、组织病理学,评估该人源化肝细胞嵌合小鼠的效果。结果诱导组小鼠血清中uPA、丙氨酸氨基转移酶(ALT)均高于未诱导组和空白对照组(P<0.01);移植组人白蛋白含量显著高于未移植组(P<0.01)。组织病理学HE染色发现人肝癌细胞参与小鼠肝组织的修复,激光共聚焦显微镜观察显示鼠肝脏中有人肝癌细胞聚集并表达人白蛋白,组化染色人角质蛋白(CK18)表达成阳性。结论新构建的转基因小鼠能通过强力霉素诱导小鼠肝脏中uPA表达,造成肝脏亚急性损伤,并通过人源肝癌细胞移植修复进而构建人源化肝细胞嵌合小鼠。

水汽造成的光模块接收光功率波动失效模式

光模块的接收光功率波动是一种常见的失效现象。对一次生产常温测试中接收光功率波动造成的模块闪断故障进行分析,经过一系列试验验证,发现该故障的失效模式与以往常见的不同,并非由器件或软件故障造成,而是由水汽造成的光路异常导致。该失效模式的发现,揭示了一种新的ROSA适配器设计风险,对研发设计和可靠性验证有重要指导意义。

立体脑电图在198例颞岛叶癫痫术前评估中的应用

目的探讨立体脑电图(SEEG)在颞岛叶药物难治性癫痫患者诊疗过程中的安全性和有效性。方法回顾性分析2014年9月—2020年12月在广东三九脑科医院癫痫中心进行SEEG置入电极患者的临床资料。所有患者均经过术前详细无创评估(包括记录发作期头皮脑电图、颅脑核磁共振成像、发作间歇期PET-CT、神经心理评估等),使用ROSA机器人置入颅内电极,监测发作间期及发作期脑电图,记录惯常发作,根据发作期颅内电极脑电图结果明确癫痫灶起始及确定功能区位置后进行手术切除。结果共564例患者接受SEEG电极置入术,其中成人414例,儿童150例。置入电极总数为6190根,其中颞、岛叶SEEG置入患者198例,共置入电极2147根。颞、岛叶患者最小8岁,最大52岁,平均(20.2±8.5)岁;男性138例(69.7%),女性60例(30.3%),平均(10.8±0.7)根/例;单侧置入141例(71.2%),双侧置入57例(28.8%),其中2例补充电极(共7根)。有3例患者出现血肿,2例行急诊手术治疗,4例患者出现少量蛛网膜下腔出血,未出现明显临床症状,2例患者因癫痫发作后出现电极折断一根。无一例出现死亡、感染、脑脊液漏等。电极最大误差为5.2 mm,最小误差为0,平均误差在2.1 mm。196例颞、岛叶癫痫患者行手术切除,2例未手术,术后随访时间6~75个月。EngelⅠ级152例(78.7%)、Ⅱ级27例(14%)、Ⅲ级9例(4.7%)、Ⅳ级5例(2.6%),其中3例失访。结论立体脑电图是一种有效和相对安全的微创手术,对涉及血管丰富及部分结构位置深在的颞叶内侧、岛叶难治性癫痫具有重要诊疗价值,熟练掌握置入技术可以减少术后并发症的发生。

不同外源生长激素处理对两个月季品种扦插生根率的影响

以不同浓度配比的萘乙酸(NAA)和吲哚丁酸(IBA)溶液处理月季(Rosa hybrid Min)“安吉拉”和月季“韧月”插穗,并对扦插后插穗的生根能力,及扦插过程中的插穗酶活特性进行分析,探究了月季生根效果最佳的激素种类及浓度配比.结果表明:当处理质量浓度为IAA 200 mg∙mL^(-1),NAA 100 mg∙mL^(-1)时,对“安吉拉”地下根部生长和地上部腋芽萌发的促进作用最明显;单独IAA 200 mg∙mL^(-1)时,对其地下根部生长的促进效果次之,但对其地上部腋芽萌发也有良好的促进效果;当IAA,NAA质量浓度都为200 mg∙mL^(-1)时,能促进其愈伤组织的诱导,但生根速度较为缓慢.

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.