Objective In this study,exogenous hormones combined with variable temperature stratification were used to relieve seed dormannia of Coptis chinensis and promote seed germination,and the effects of gibberellin on the physiological,biochemical and embryonic morphological changes of C.chinensis seeds during post-mature germination were further evaluated.Methods Using orthogonal test,different exogenous hormones(GA3,6-BA,NAA,ABA)and different temperature changing stratification(20℃/4℃)were set up,and the germination index(germination rate and split rate)of C.chinensis seeds were statistically compared.The activity of antioxidant enzymes(peroxidase POD,catalase CAT),the content of malondialdehyde(MDA)and endogenous hormones(GA,IAA,ABA)during the germination of C.chinensis seeds under optimal treatment were measured.The microscopic structure of the seeds was observed by paraffin embedding section.Results The results of range analysis showed that the best budding method was:100 mg·L-1 of gibberellin for 12 h,20℃for 60 day,4℃for 10 day.Physiological and biochemical index analysis showed that 100 mg·L-1 exogenous gibberellin treatment promoted the endogenous GA content to reach the peak in advance,maintained the POD and CAT contents at a higher level,and maintained the malondialdehyde(MDA)content at a lower level.The transformation from torpedo embryo to cotyledonous embryo was observed by electron microscope during postmature germination.Conclusion Exogenous gibberellin combined with variable temperature stratification treatment could significantly accelerate seed germination of C.chinensis,mainly by promoting seed embryo morphological development,affecting the contents of GA,POD and CAT,and rapidly relieving seed dormancy to promote seed germination.Compared with natural conditions,the germination period could be shortened by about six months.

Objective Systematic identification and analysis of the cytochrome P450(CYP450)gene family members and their functions in Chrysanthemum indicum L.to provide a basis for further studies on their roles in the flavonoid synthesis pathway.Methods Based on the genomic data of a diploid Chrysanthemum indicum L.,the members of the P450 gene family were identified and analyzed using bioinformatics methods.Moreover,the transcriptome and quasi-targeted metabolome analysis were combined to screen the flavone synthase Ⅱ(FNSII)genes from the CYP93 family,and their expression levels were tested in different tissues of C.indicum using real-time fluorescence quantitative PCR(qPCR).Results A total of 460 P450 genes were identified from the C.indicum genome,belonging to 43 families within 8 family clans.The encoded proteins ranged from 336 to 1538 amino acids in length,with a relative molecular mass between 38.01 to 175.00 kDa,and an isoelectric point between 5.61 to 9.71.Chromosome localization analysis indicated that these 460 P450 genes were unevenly distributed on nine chromosomes,with the highest number of light-responsive elements in the promoter regions.Expression pattern analysis showed that the expression levels of two FNSII genes(CindChr2G00102820.1 and CindChr8G00552890.1)were significantly higher in leaf of C.indicum than in flower.Conclusion The comprehensive analysis of the P450 gene family in C.indicum is helpful to further elucidate the function of the P450 gene family in flavonoid biosynthesis.

Objective Five chloroplast(cp)genomes from members of genus Polygonatum were assembled by hybrid assembly technique,and their intraspecic and interspecific differences were analyzed by comparative genomic method.Codon usage patterns and influencing factors were determined,and the cp genome data were applied to understand the phylogenomic relationships in the entire genus Polygonatum along with the available data.Methods In this study,the chloroplast genomes of 5 species of genus Polygonatum were assembled using Unicycler software.Sequence alignment,collinearity analysis,boundary analysis and other methods were used to evaluate the interspecific differences of these five species.Nucleotide polymorphism analysis was used to discover the high-variation sites of the five species and their related species,predict the distribution of different long repeat sequences and SSRs,and then analyze the use bias of Polygonatum code.Finally,phylogenetic tree was constructed with the coding sequences of other 47 genus Polygonatum and their closely related species to explore their phylogenetic relationships in this study.Results ①Chloroplast genomes of 155 408-155 623 bp were assembled from five species of Polygonatum.A total of 132-133 genes were annotated,and 369 long repeats and 1553 simple repeats were detected.②The contraction and expansion of chloroplast genomes in 8 species were not obvious at the IRs boundary,and the size and distribution of individual genes at the LSC-IRs-SSC boundary,such as ndhF gene and ycf1 gene,were slightly different.No interspecific or intraspecific rearrangement was observed in 8 species.③ The high-variation regions of the 8 chloroplast genomes are mainly located in two single-copy regions,the duplicate copy region is relatively conserved,and the coding region is more conserved than the non-coding region.High nucleotide polymorphic loci rps16-trnQ,trnS-trnG,trnTUGU-trnL,ndhF-rpl32 and rpl32-trnL are located in the single copy region and most of them are gene spacer regions.④ The codon preference results showed that the codon preference of the five species was similar and mainly affected by selection pressure,and the third base of the codon played A dominant role and mainly ended in A/U.RSCU clustering heat map shows that PK and PZ,PCB and PS have close relationship.⑤ Phylogenetic trees divided 52 species into five branches:Ⅰ,Ⅱ,Ⅲ,ⅣandⅤ.PS,PK,PCB,PCZ and PZ were divided into ⅣandⅤbranches,among which PK and PZ were most closely related,while PCZ was more distant than the other four,was divided into the Ⅴbranch alone.Conclusion This study provided a reference for the phylogenetic research and molecular marker development of the medicinal plants of the Polygonum genus.

Objective The three UGT genes involved in the synthesis of flavonoid glycosides in the wild chrysanthemum were cloned,fused and expressed in prokaryotic system,which provided a basis for further study of the functions of these genes in the synthesis of flavonoid compounds.Methods three candidate UDP-glycosyltransferase(UGT)family genes were successfully cloned from flower tissue cDNA of diploid Chrysanthemum indicum.L in Hubei,namely:CiUGT1(1425 bp),CiUGT2(1311 bp)and CiUGT3(1314 bp)encoded opal fraction of 51.72,48.08 and 48.28 kDa,respectively.After the physical and chemical properties of the protein were analyzed,the recombinant plasmid was constructed by seamless Klon technology,and the recombinant proteins of CiUGT2 and CiUGT3 were successfully expressed in Escherichia coli.Results SDS-PAGE analysis showed that the purified CiUGT2 and CiUGT3 protein samples were about 0.2 mg/mL and 1.4 mg/mL,respectively.Conclusion Two candidate UGTs were obtained to catalyze the synthesis of flavonoid 7-O-glucosides in Chrysanthemum indicum.L,which provide the theoretical basis for the molecular mechanism of flavonoid glycosides biosynthesis in the capitulum of Chrysanthemum indicum.L.

Objective Based on the transcriptome data of Scolopendra subspinipes mutilans L.Koch,SSR primers were designed and developed to evaluate the genetic diversity and analyze the population structure of the germplasm resources of S.mutilans,which can provide a theoretical basis for the development and utilization of the germplasm resources of S.mutilans.Methods We collected 301 samples of S.mutilans from major production areas,and analyzed the population genetic diversity and population structure of the germplasm resources of S.mutilans using 18 pairs of highly polymorphic SSR primers after screening.Results A total of 142 alleles were amplified by 18 primer pairs across all samples,with an average of 7.89 alleles amplified per locus.The mean of observed heterozygosity(HO)of the 18 populations was 0.39,the mean of expected heterozygosity(HE)was 0.46,the mean of polymorphism information content(PIC)was 0.41,the mean of Nei's gene diversity index(H)was 0.46,and the mean of Shannon information index(I)was 0.81.The results showed that the wild resources of S.mutilans collected in this study had a high level of genetic diversity,Four populations in Anhui and Zhejiang had higher genetic diversity than populations in Hubei.Molecular analysis of variance(AMOVA)of the population showed that most of the genetic variation in the materials of this study mainly occurred within populations.The STRUCTURE analysis of germplasm resources showed that all germplasm could be divided into four populations.Conclusion This study showed that the natural population of S.mutilans possesses high genetic diversity with high genetic variation and is better adapted to changes in the environment.Our results provided a research basis for the rational utilization of the germplasm resources of S.mutilans and the selection and breeding of superior varieties.

Objective To provide scientific basis for layout planning and standardized cultivation the species of Chrysanthemum indicum complex in future periods,the potential ecological suitable areas was analyzed,.Methods By collecting distribution data of 1379 valid C.indicum complex species nationwide and 104 ecological factors,combined with MaxEnt model and ArcGIS software,the potential ecological distribution areas of C.indicum complex under 17 different climate scenarios was analyzed.Results Precipitation and solar radiation are key environmental variables affecting the suitable habitats of C.indicum complex.The most suitable regions for the growth of C.indicum complex was concentrated in Hubei(C.indicum L.),Shanxi(C.lavandulifolium),Jiangsu(C.nankingense),and Hubei(C.indicum var.aromaticum)regions.Under different scenarios,the suitable habitat area for C.indicum L.and C.lavandulifolium showed an overall decreasing trend,while those for C.indicum var.aromaticum and C.nankingense showed an expanding trend.The centroid shift indicates sensitivity of C.indicum complex to climate change.Conclusion The potential distribution areas of C.indicum complex is classified into ecological suitability levels,providing important reference for the sustainable use,introduction cultivation,and scientific zoning of related resources.

Objective To investigate the mechanism of lobetyolin's intervention in HepG2 cells abnormal cholesterol metabolism.Methods This study used oleic acid(OA)stimulation of HepG2 cells as a model.MTT assay,oil red O staining,biochemical kit assay,qRT-PCR assay,Western blot assay and NBD labeled cholesterol effection assay were used to study the effect of lobetyolin on cholesterol metabolism in HepG2 cells.Results The results showed that lobetyolin could reduce the content of lipid drops,the levels of triglyceride(TG)and total cholesterol(TC)in HepG2 cells stimulated by OA,increase cholesterol effection rate,and up-regulate cytochrome 7A1(CYP7A1),liver x receptor α(LXRα),ATP-binding cassette transporter A1(ABCA1),peroxisome proliferator-activated receptor(PPARγ)and other mRNA or protein expression levels.However,combined intervention with PPARγ antagonist Mifobate(SR-202)can significantly inhibit the promoting effect of lobetyolin on cholesterol metabolism in HepG2 cells.Conclusion This study revealed that lobetyolin can improve the cholesterol effection rate of HepG2 cells stimulated by OA and promote cholesterol catabolism,and the mechanism of action may be related to the regulation of PPARγ/CYP7A1 pathway.

Background When discussing the new concept of"pancreas spleen integration"in the early stage,we proposed that dampness heat stagnation in the spleen and deficiency due to stagnation are the traditional Chinese medicine(TCM)pathogenesis elements of pancreatitis-induced pancreatic ductal adenocarcinoma,among which dampness heat is the soil of pancreatitis deteriorates into pancreatic cancer.Objective Based on the TCM pathogenesis elements of pancreatitis to cancer with"damp heat"as the main factor,data mining technology was used to analyze the properties,channel conversion and active ingredients of heat-clearing Chinese medicines with anti-inflammatory and anti-tumor effects,and to summarize the drug characteristics of these Chinese medicines.To prospectively predict the use of drugs in the transformation process of pancreatitis to pancreatic cancer.Methods Taking Traditional Chinese Medicine(10th Edition of China Traditional Chinese Medicine Press)as the drug data source,66 heat-clearing herbs were searched in the literature database of CNKI and PubMed of inflammation * TCM or(inflammation+tumor)* TCM.The active ingredients were analyzed in the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP).Results Among the 66 heat-clearing drugs(excluding adjunct drugs),58 herbs had anti-inflammatory effects,53 herbs had anti-tumor effects,and 48 herbs had both anti-inflammatory and anti-tumor effects.The three groups of drugs were mainly bitter,sweet and pungent in five flavors,and were mainly liver,stomach,lung,heart and large intestine in meridian affinity.Five flavors and meridian affinity were highly similar.In the TCMSP,the"five principles of drug class"were used to screen the active ingredients of anti-inflammatory and anti-tumor drugs.A total of 1041 active ingredient data were screened,and 798 active ingredient data were screened after duplicate items were deleted.Top three active ingredients were Luteolin,Kaempferol and Acacetin.Conclusion Based on the analysis of five flavors,meridian affinity and active ingredients of heat-clearing medicine,this study found that the anti-inflammatory and anti-tumor effects of these Chinese herbs are highly overlapping,it has important guiding significance for the drug research and clinical prescription of pancreatitis-induced pancreatic ductal adenocarcinoma.

Objective Bibliometric analysis software VOSviewer and CiteSpace were applied to analyze literatures related to acupuncture for myocardial Ischemia/Reperfusion(I/R)injury published in Chinese and English,to explore the status quo,dynamic trend and hot spots in the field,in order to provide reference for future research.Methods Based on the literature on acupuncture and moxibustion for myocardial I/R injury published in CNKI and Web of Science Core Collection database,VOSviewer and CiteSpace software were used to analyze literature publication trends,authors,institutions,countries,journals,keywords and citations,and draw graphs and tables to show the development status of the field and analyze the future development direction.Results Both Chinese and English literatures on the treatment of myocardial I/R injury by acupuncture and moxibustion show an increasing trend,which is one of the academic hotspots with great potential at present.In terms of Chinese literature,Yan Jie is the scholar who has published the most papers,and Hunan University of Traditional Chinese Medicine is the representative of the high production institution.The journal of Acupuncture Research has published the most papers.In terms of English literature,lu sheng-feng is the representative scholar with a high number of publications,Nanjing University of Chinese Medicine is the institution with the largest number of publications,and Evidence-Based Complementary and Alternative Medicine is the journal with the largest number of publications.The authors in the field have weak interinstitutional cooperation and exchange,and their research is mainly from China,with little international collaboration and exchange.At present,acupuncture treatment of myocardial I/R injury covers three aspects:specific acupoints,molecular biological mechanisms and clinical studies,which mainly focus on seven hot spots.Conclusion The bibliometric visual analysis visually demonstrated the current status,hot spots and frontier trends of acupuncture in the treatment of myocardial I/R injury.In the future,it is necessary to strengthen international collaboration,promote exchanges and cooperation between scholars and institutions,and promote the publication of multi-center,large-sample and high-level clinical randomized controlled studies;At the same time,the effect mechanism was further discussed from the perspectives of new molecular biological mechanisms,new intervention measures and point selection,and clinical translational research was carried out.

Objective To investigate the improvement of interleukin-1 receptor associated kinase(IRAK1)/nuclear factor κB(NF-κB)signaling pathway regulated by"srperficiality-origin matching acupoints"electroacupuncture on intestinal damage in diabetic rats and lipid metabolism.Methods SD rats were randomly separated into control group,diabetes group,diabetes+electroacupuncture group,diabetes+pcDNA group,diabetes+pcDNA-IRAK1 group,diabetes+electroacupuncture+pcDNA group,and diabetes+electroacupuncture+pcDNA-IRAK1 group,18 rats in each group.Except for the control group,rats in other groups were fed with high glucose and high fat diet and intraperitoneal injection of streptozotocin(STZ)to construct the model of diabetes rats.After successful modeling,rats in each group were treated every other day for 8 consecutive weeks.The weight,fasting blood glucose,serum triglycerides and total cholesterol levels of rats were detected.16S rRNA amplicon sequencing was applied to detect the relative abundance of rat gut microbiota;the intestinal permeability and water content of rats were tested.Immunohistochemistry was applied to detect the average optical densities of zonula occludens-1(ZO-1)proteins in small intestine tissue.Western blot was applied to detect the expression of IRAK1,phosphorylated nuclear factor inhibitory protein(p-IκBα),and phosphorylated nuclear factor B p65(p-NF-κB p65)proteins in rat small intestine tissue.Results Compared with the control group,the levels of fasting blood glucose,triglyceride,total cholesterol,relative abundances of Firmicutes,Proteobacteria,Actinomycetes,Allobaculum,fluorescein isothiocyanate(FITC)-glucan concentration,intestinal water content and the expression of IRAK1,p-IκBα,p-NF-κB p65 proteins in the diabetes group increased significantly(P<0.05).Weight,the relative abundance of Bacteroidetes,Lactobacillus,Romboutsia,Lachnospiraceae,Ruminocoaceae,the average optical densities of occludin and ZO-1 proteins decreased significantly(P<0.05).Compared with the diabetes group,the levels of fasting blood glucose,triglyceride,total cholesterol,relative abundances of Firmicutes,Proteobacteria,Actinomycetes,Allobaculum,FITC-glucan concentration,intestinal water content and the expression of IRAK1,p-IκBα,p-NF-κB p65 proteins in the diabetes+electroacupuncture group decreased significantly(P<0.05).Weight,the relative abundance of Bacteroidetes,Lactobacillus,Romboutsia,Lachnospiraceae,Ruminocoaceae,the average optical densities of occludin and ZO-1 proteins increased significantly(P<0.05),the change trend of corresponding indicators in the diabetes+pcDNA-IRAK1 group was opposite to the above(P<0.05).Compared with the diabetes+electroacupuncture group and the diabetes+electroacupuncture+pcDNA group,the levels of fasting blood glucose,triglyceride,total cholesterol,relative abundances of Firmicutes,Proteobacteria,Actinomycetes,Allobaculum,FITC-glucan concentration,intestinal water content and the expression of IRAK1,p-IκBα,p-NF-κB p65 proteins in the diabetes+electroacupuncture+pcDNA-IRAK1 group increased significantly(P<0.05).Weight,the relative abundance of Bacteroidetes,Lactobacillus,Romboutsia,Lachnospiraceae,Ruminocoaceae,the average optical densities of occludin and ZO-1 proteins decreased significantly(P<0.05).Conclusion"Srperficiality-origin matching acupoints"electroacupuncture may improve intestinal flora imbalance and intestinal barrier damage in diabetes rats by inhibiting IRAK1/NF-κB signaling pathway,and improve diabetic glucose and lipid metabolism.