To classify and identified the nematode parasized in ruminant in Qinghai province,the mature nematode specimen parasized in Tibetan sheep,goat,yak,ox and camel was collected and classified systematically,by adopting classfication systems of Yamaguti(1961).All those specimens belong to 24 genera of 15 families of 6 orders in nematade of Nemathelminthes Class,of which 85 kinds of nematodes were sorted out,among them 13 lung nematodes,71 digestine nematode and 1 muscle nematode were classified.

Objective To isolate endophytic fungi from the rare medicinal plants ephedra, detect the production of ephedrine alkaloids, and explore the optimum fermentation conditions.Methods The endophytic fungi were isolated from the herbaceous stems treated.The ephedrine alkaloids were preliminarily detected by Dragendorff's reagent.The fungi strains producing ephedrine alkaloids were screened by HPLC method with reference substance of ephedrine, pseudoephedrine and methephedrine.After the optimum carbon source and nitrogen source were determined with total ephedrine alkaloids as evaluation indicators by HPLC, the orthogonal test of L16 (45) was designed to investigate five levels of carbon source amount, nitrogen source amount, pH value, temperature, fermentation time.Results The ephedra herbaceous stems endophytic fungus-Es2 mycelium extract after fermentation contained ephedrine, pseudoephedrine and methephedrine.Es3 mycelium extract containing pseudoephedrine after fermentation.Lactose and ammonium sulfate were as the most suitable carbon source and nitrogen source of Es2 fermentation to produce ephedrine alkaloids.Es produced the highest yield of ephedra alkaloids when the carbon source 30 g/L, nitrogen source 4 g/L, pH5.0, temperature 30℃, fermented for 8 days.Conclusion The ephedrine alkaloids of industrial production could be realized by microorganism fermentation method, which has very important significance to alleviate the ephedrine market demand pressure on the environment.

Objective To study the genetic diversity of Fritillaria thunbergii,a traditional Chinese herb in Zhejiang Province in China.Methods The genetic diversity of six representational populations of F.thunbergii including 32 individuals was investigated by amplified fragment length polymorphism(AFLP) maker technique.Results The genetic diversity was revealed as follow: the Nei′s genetic diversity index(He) 0.169 0?0.175 7,Shannon′s information index(I) 0.269 8?0.245 3,percentage of polymorphic loci(PPB) was 76.85% at the species level;Ht 0.169 0?0.030 9,and Hs 0.150 8?0.024 0,I 0.233 3?0.261 9, PPB was 50.38% at population level.The genetic differentiation index(Gst) was 0.107 6,Nm 4.147 0.The result of dendrogram of six populations indicated that Dongyang and Yongkang populations shared the minimum genetic distance(0.015 0),they were classified into a group,and Xiangshan and Jinyun populations shared the maximum genetic distance(0.032 4).Conclusion The genetic diversity of F.thunbergii cultivated in Zhejiang Province is very rich,which could ensure the long-term survival of F.thunbergii.But the genetic diversity of F.thunbergii is relatively higher in population levels while lower at the species levels and the degree of genetic differentiation occured among the populations is not significant.The germplasm resources are relatively stable among these six populations.These populations could be used to breed the fine strains of F.thunbergii as the bases.

Objective To observe the genetic structure of cultivated Scrophularia ningpoensis in Zhejiang Province.Methods The genetic structures of six typical S.ningpoensis populations were analyzed by fluorescence AFLP marker.Results Bands(12 552) were generated by seven pairs of AFLP primer combinations,of which 8 808 were polymorphic,and the polymorphic rate was 70.17%.The variety ranges of PPB among different populations were 41.67%—55.56%,and 47.30% in average.I was between 0.190 8—0.238 3,and 0.221 8 in average.Ne was between 1.201 4—1.280 6,and 1.236 9 in average.Gst was 0.127 1,Nm was 3.432 4.UPGMA Cluster analysis showed that the six populations can be divided into two clusters,as that of Tiantai,Jinyun,and Jingning were one sub-cluster,and Dongyang,Pan′an,and Xianju were another one sub-cluster.Conclusion There is a relative high genetic diversity level in cultured S.ningpoensis of Zhejiang Province.Genetic differentiation exists among populations,but it exists in population mostly.There is a relative high genetic intercommunion among populations.The genetic distance is not related to the geographic environment.

Medicinal plant germplasm resources are the foundation of the modern development of traditional Chinese medicine. In-depth study of medicinal plant germplasm resources is a prerequisite for cultivating fine varieties and ensuring the output and standard quality of traditional Chinese medicine（TCM）. Traditional identification methods start with appearance and are greatly affected by natural environment and human factors，with a low efficiency and accuracy of identification are generally low molecularin general. Due to such advantages as easy operation，high sensitivity，accurate results， molecular biology technology has been widely used in the related research of relevant studies for medicinal plant germplasm resources due to its advantages of easy operation，high sensitivity，accurate results，etc. It mainly involving the distinction between wild and cultivated products，researchstudy on substitutes of TCM，identification of Chinese patent medicine，good variety marker breeding，genetic diversity researchstudy，genetic map establishment and omics research，etcstudy. Among them，omics researchstudy is divided into genomics，transcriptomics，metabolomics，and proteomics due toby different analysis purposes. Genomics is divided into three sub-fields namely structural genomics，functional genomics， and comparative genomics. Eukaryotes Because eukaryotes have nuclei and organelles，so omics researchstudy also includes chloroplast genomics，mitochondrial genomics，nuclear genomics，and plastid genomics. Among them，the chloroplast genome has a simple structure，small molecular weight，and good conservation，while the mitochondrial genome has a strong variability and complex structure，the nuclear genome data isfeatures complex， data and the nucleus contains no ribosomes in nucleus，resulting in spatiotemporal differences in the translation process，even if repeated repeatedly test， the result of and the test is alsoresults remained uncertain， even after repeated tests. The molecular biology technology and omics researchstudy involved in theby current medicinal plant researchstudy still hashave shortcomings，and there iswith a large room for development，which needs and need further improvement and supplementation. This articlepaper successively introduces the characteristics and applications of cytology，molecular markers，and omics researchstudy techniques in the identification of medicinal germplasm resources，providingin order to provide a reference for subsequent identification，development and utilization of medicinal plant germplasm resources.

OBJECTIVES: To explore the mRNA differential expressions and the sequential change pattern in acute myocardial infarction (AMI) mice. METHODS: The AMI mice relevant dataset GSE4648 was downloaded from Gene Expression Omnibus (GEO). In the dataset, 6 left ventricular myocardial tissue samples were selected at 0.25, 1, 4, 12, 24 and 48 h after operation in AMI group and sham control group, and 6 left ventricular myocardial tissue samples were selected in blank control group, a total of 78 samples were analyzed. Differentially expressed genes (DEGs) were analyzed by R/Bioconductor package limma, functional pathway enrichment analysis was performed by clusterProfiler, protein-protein interaction (PPI) network was constructed by STRING database and Cytoscape software, the key genes were identified by Degree topological algorithm, cluster sequential changes on DEGs were analyzed by Mfuzz. RESULTS: A total of 1 320 DEGs were associated with the development of AMI. Functional enrichment results included cellular catabolic process, regulation of inflammatory response, development of muscle system and vasculature system, cell adhesion and signaling pathways mainly enriched in mitogen-activated protein kinase (MAPK) signaling pathway. The key genes of AMI included MYL7, TSC22D2, HSPA1A, BTG2, NR4A1, RYR2 were up-regulated or down-regulated at 0.25-48 h after the occurrence of AMI. CONCLUSIONS The functional signaling pathway of DEGs and the sequential expression of key genes in AMI may provide a reference for the forensic identification of AMI.
Animals ; Computational Biology/methods* ; Gene Expression Profiling/methods* ; Mice ; Mitogen-Activated Protein Kinases/metabolism* ; Myocardial Infarction/metabolism* ; RNA, Messenger ; Ryanodine Receptor Calcium Release Channel/metabolism* ; Transcriptome

OBJECTIVES: To explore the differential expression of messenger RNA (mRNA) in myocardial tissues of rats with sudden coronary death (SCD), and to provide ideas for the forensic identification of SCD. METHODS: The rat SCD model was established, and the transcriptome sequencing was performed by next-generation sequencing technology. Differentially expressed genes (DEGs) in myocardial tissues of SCD rats were screened by using the R package limma. A protein-protein interaction (PPI) network was constructed by using the STRING database and Cytoscape 3.8.2 on DEG, and hub genes were screened based on cytoHubba plug-in. Finally, the R package clusterProfiler was used to analyze the biological function and signal pathway enrichment of the selected DEG. RESULTS: A total of 177 DEGs were associated with SCD and were mainly involved in the renin-angiotensin system and PI3K-Akt signaling pathway. The genes including angiotensinogen (AGT), complement component 4a (C4a), Fos proto-oncogene (FOS) and others played key roles in the development of SCD. CONCLUSIONS Genes such as AGT, C4a, FOS and other genes are expected to be potential biomarkers for forensic identification of SCD. The study based on mRNA expression profile can provide a reference for forensic identification of SCD.
Rats ; Animals ; RNA, Messenger/genetics* ; Gene Regulatory Networks ; Gene Expression Profiling ; Phosphatidylinositol 3-Kinases/genetics* ; Biomarkers

Objective To study the DNA methylation of nucleated cells in peripheral blood of patients died from anaphylactic shock caused by cephalosporin drugs and to provide a new research direction and basis for the forensic diagnosis of shock caused by drug hypersensitiveness. Methods Methylation microarray was used to detect DNA methylation of nucleated cells in peripheral blood of patients died from anaphylactic shock caused by cephalosporin drugs and normal subjects. Sequencing data and chip data were analyzed for differences in DNA methylation using R language methylkit, ChAMP package. Random forest algorithm was used to evaluate the importance of the DNA methylation differential sites. Results Differential sites of DNA methylation highly associated with anaphylaxis caused by cephalosporin drugs were obtained at loci such as ETS1, PRR23B and GNAS. Conclusion Cephalosporin allergy is associated with DNA methylation, and DNA methylation may be a new strategy for forensic identification of anaphylactic shock and death.
Anaphylaxis/genetics* ; DNA Methylation ; Forensic Medicine ; Humans

In this study, RAW264.7 cells were employed to investigate the effects of honey-processed Astragalus on their energy metabolism and polarization, and explore the scientific connotation of the enhanced efficacy of honey-processed Astragalus on invigorating spleen-stomach and replenishing Qi. The medicated sera were prepared by intragastric administration of rats with water extracts of crude and honey-processed Astragalus, and the composition changes of medicated sera of crude and honey-processed Astragalus were analyzed by using LC-MS technology. The cell survival rates were detected and the concentrations of medicated sera were screened through CCK-8 assay. The differences of cell phagocytic rates, ATP energy metabolism, and NO secretion between crude and honey-processed Astragalus were evaluated by using neutral red phagocytosis assay, ATP detection kit, and NO detection kit. The effects of crude and honey-processed Astragalus on TNF-α secretion of RAW264.7 cells were detected by employing ELISA kit. The effects of crude and honey-processed Astragalus on polarization of RAW264.7 cells were evaluated by utilizing flow cytometry. The differential metabolites related to glycolysis in cell lysates and culture media were screened by using LC-MS technology. The experiment was approved by the experimental animal ethics committee from Shanxi University of Chinese Medicine (No. AWE202407352). The results showed that the contents of betaine, amino acids, and ononin in the prepared medicated sera of rats treated by intragastric administration with water extracts of honey-processed Astragalus increased compared to those in crude one. The results of CCK-8 experiment showed that there were no cytotoxic effects on RAW264.7 cells in the medicated sera of crude and honey-processd Astragalus at different concentration. The phagocytic index and ATP yield both increased to varying degrees after administration of the medicated sera to cells. The secretion of NO in normal and inflammatory cells increased and decreased respectively, and the effect of honey-processed Astragalus was better than that of crude one. The results of ELISA kit showed that the medicated sera of both crude and honey-processed Astragalus could promote the secretion of TNF-α in a concentration-dependent manner, and the promoting effect of honey-processed Astragalus was stronger than that of crude one. The results of flow cytometry showed that the medicated sera of both crude and honey-processed Astragalus could promote M1-type polarization and inhibit M2-type polarization of RAW264.7 cells, and the effect of honey-processed Astragalus was better than that of crude one. Compared to crude Astragalus, the metabolites related to glycolysis in the cell lysates and culture media of the medicated sera of honey-processed Astragalus were generally on the rise, indicating that the effect on promoting glycolysis of honey-processed Astragalus was better than that of crude one. In summary, honey-processed Astragalus can promote the polarization and energy metabolism of RAW264.7 cells, and participate in positive immune regulation, which is correlated with its enhanced effect of invigorating spleen-stomach and replenishing Qi.

ObjectiveTo analyze the differential components in water extract of Chuanxiong Rhizoma before and after processing with wine， and to explore the molecular mechanism of Chuanxiong Rhizoma processed with wine in enhancing anti-cerebral ischemia injury. MethodUltra high performance liquid chromatography tandem quadrupole orbitrap high resolution mass spectrometry （UHPLC-Q-Orbitrap HRMS） was used to qualitatively analyze the main chemical components in water extract of Chuanxiong Rhizoma based on the spectral information of compound， comparison of reference substance and references. The chemical pattern recognition method was used to screen the differential components of Chuanxiong Rhizoma before and after processing. Based on these differential components， the potential targets of differential components were predicted by online databases， and the related targets of cerebral ischemia were searched. Cytoscape 3.6.0 was used to establish the network diagram of differential components-action targets-diseases of Chuanxiong Rhizoma processed with wine. The protein-protein interaction （PPI） network of intersection targets was constructed by STRING 11.5. The potential targets of differential components against cerebral ischemia were analyzed by Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis through DAVID 6.8. At the same time， the chemical compounds with high relative content and increased peak area after wine processing were docked with their corresponding targets to verify the mechanism of enhanced effect after wine processing. ResultA total of 71 chemical components were identified from Chuanxiong Rhizoma， 34 differential components and 603 potential targets were screened out. At the same time， a total of 769 disease targets and 60 intersection targets were obtained. Seven key targets were identified through PPI network analysis， including JUN， signal transducer and activator of transcription 3 （STAT3）， mitogen-activated protein kinase 3 （MAPK3）， interleukin-1β （IL-1β）， vascular endothelial growth factor A （VEGFA）， Caspase-3 （CASP3） and mtrix metalloproteinase 9 （MMP9）. Tumor necrosis factor （TNF） signaling pathway was the main differential signaling pathway. The results of molecular docking showed that differential components （senkyunolide K， senkyunolide F， 3-n-butylphthalide， Z，Z′-6，8′，7，3′-diligustilide， ferulic acid and Z-ligustilide） and corresponding targets had good binding activities. ConclusionThe synergistic mechanism of Chuanxiong Rhizoma processed with wine may be related to the enhanced inhibitory effect of inflammatory reaction.