A novel biological sources consistency evaluation method reveals high level of biodiversity within wild natural medicine:A case study of Amynthas earthworms as"Guang Dilong"

For wild natural medicine,unanticipated biodiversity as species or varieties with similar morphological characteristics and sympatric distribution may co-exist in a single batch of medical materials,which affects the efficacy and safety of clinical medication.DNA barcoding as an effective species identification tool is limited by its low sample throughput nature.In this study,combining DNA minibarcode,DNA metabarcoding and species delimitation method,a novel biological sources consistency evaluation strategy was proposed,and high level of interspecific and intraspecific variations were observed and validated among 5376 Amynthas samples from 19 sampling points regarded as"Guang Dilong"and 25 batches of proprietary Chinese medicines.Besides Amynthas aspergillum as the authentic source,8 other Molecular Operational Taxonomic Units(MOTUs)were elucidated.Significantly,even the subgroups within A.aspergillum revealed here differ significantly on chemical compositions and biological activity.Fortunately,this biodiversity could be controlled when the collection was limited to designated areas,as proved by 2796"decoction pieces"samples.This batch biological identification method should be introduced as a novel concept regarding natural medicine quality control,and to offer guidelines for in-situ conservation and breeding bases construction of wild natural medicine.

Combination of Shuxuetong and aspirin protects against cerebral ischemia/reperfusion injury through ameliorating coagulation and fibrinolysis system

Objective：To explore the influence of the combination of Shuxuetong（SXT）and aspirin on coagulation and fibrinolytic system of rats.Methods：Suture method was applied to establish focal cerebral ischemia-reperfusion injury models in rats.SD rats were randomly divided into sham group,middle cerebral artery occlusion/reperfusion（MCAO/R）group,aspirin group,SXT group,and SXT＋aspirin group（S＆A）.The neurological deficits were assessed according to Longa’s grade 5 scoring method.The cerebral edema was detected by measuring the content of water in brain tissue.The volume of cerebral infarction was observed by 2,3,5-Triphenyltetrazolium chloride（TTC）staining.Blood plasma was collected by abdominal aortic method to test maximum platelet aggregation rate and four blood coagulation.CD61,CD62p,6-keto prostaglandin F1αantithrombinⅢ（AT-Ⅲ）,D-dimer,plasminogen activator inhibitor-1（PAI-1）,tissue factor（TF）,tissue plasminogen activator（t-PA）,platelet thromboxaneand von Willebrand factor（v WF）content in rat plasma were detected by ELISA.Results：SXT combined with aspirin could improve the neurological deficits,alleviate cerebral edema,and decrease the cerebral infarct value.Compared with the sham operation group,fibrinogen（FIB）,6-AT-III,and t-PA in model group were significantly decreased;Compared with the model group,the above-mentioned indexes in SXT and aspirin treatment group were significantly increased.The prothrombin time（PT）,activated partial thromboplastin time（APTT）,thrombin time（TT）,D-dimer,PAI-1,TF,TXB2,and v WF of the model group were significantly increased;The above-mentioned indexes in blood SXT＋aspirin treated group were significantly decreased.There was a significant difference between the combined group and SXT group.The maximum concentration of plateletsin aspirin treated rats was significantly decreased,however,MPAR was reversed in SXT＋aspirin treated group.Conclusion：SXT combined with aspirin can effectively inhibit platelet activation,regulate the maximum concentration of platelets,and improve coagulation function and fibrinolysis system.