OBJECTIVETo study testing methods of seed quality, and provide a basis for establishing seed testing specification of Cyathula officinalis.

METHODReferring to the Specifications for Agricultural Seed Testing, the optimal testing methods of seed quality of C. officinalis were screened.

RESULT AND CONCLUSIONThe testing method for C. officinalis seed quality has been initially established. At least 8 g seeds should be sampled and passed through 20-mesh sieve for purity analysis. The phenotypic observation and size measurement were used for authenticity testing. The seeds were inoculated directly on PDA medium, cultured 5 days on 28 degrees C for seed health testing. The weight of 1 000 seeds was determined by using the 500-seed method. The water content of the seeds was determined under the higher temperature (133 +/- 2) degrees C for 3 hours. The seeds were dipped into 0.1% TTC solution 3 hours for determining viability. The seeds were cultured on pleated paper at 25 degrees C for 2-9 days for germination testing.

Germination ; physiology ; Phenotype ; Plants, Medicinal ; embryology ; growth & development ; Quality Control ; Seeds ; chemistry ; Temperature ; Water ; analysis

Objective·To construct a self-healing injectable hyaluronic acid(HA)-based hydrogel(HAPD-Cu)and investigate the effects of different copper ions on the properties of the hydrogel and its vasogenic effiicacy to evaluate its feasibility for clinical wound healing.Methods·Bisphosphonated hyaluronic acid(HAPD)was prepared via a blue-light mediated thiol-ene click reaction between thiolated hyaluronic acid(HASH)and acrylated bisphosphonate(Ac-PD)in the presence of photoinitiator 2959.Then,HAPD was further interacted with Cu2+through metal coordination to prepare HAPD-Cu hydrogels with different Cu2+concentrations,i.e.HAPD-Cu1,HAPD-Cu2,HAPD-Cu3 and HAPD-Cu4.The molecular structures of HASH,Ac-PD,HAPD and HAPD-Cu were verified with 1HNMR and FTIR.Microscopic morphology of HAPD-Cu was observed under SEM.The shear-thinning and self-healing properties of HAPD-Cu were verified by rheometer.The Cu2+release from HAPD-Cu was determined with ICP.Live-dead staining and CCK-8 assay were applied to evaluate the biocompatibility of HAPD-Cu.The in vitro vasculogenic activity of HAPD-Cu was determined by a tubule-forming assay with human umbilical vein vascular endothelial cells and the in vivo vasculogenic activity of HAPD-Cu was assessed by CD31 tissue staining.A rat wound defect model was established in vitro to evaluate its actual repair effect.Results·The preparation of the materials was demonstrated through chemical qualitative and quantitative analytical means.In vitro studies showed that all HAPD-Cu with a loose porous internal structure exhibited outstanding self-healing,injectability and degradability,with a one-week degradation cycle and abrupt release behavior,which can meet the needs of wound healing cycle.All HAPD-Cu showed good biocompatibility except HAPD-Cu4,due to its high Cu2+concentrations.Moreover,its angiogenic effect in vitro or in vivo was enhanced with increasing Cu2+concentrations within the permissible Cu2+concentration range.In vitro wound model experiments also showed that the HAPD-Cu hydrogel significantly promoted wound healing compared with the control group.Conclusion·HAPD-Cu hydrogel constructed via the metal coordination shows excellent shape plasticity,allowing the filling of defective sites in a minimally invasive form,and the release of Cu2+greatly facilitates the establishment of early vascular networks,with giant potential for use in the repair of clinically irregular wounds.