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Cardiovascular and cerebrovascular diseases are the leading cause of death for residents in China. Danhong Injection(DHI) decoction piece is prepared from Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos, with the function of promoting the blood circulation, removing the blood stasis, relaxing the sinews and dredging the collaterals. In recent years, about 100 million bottles of DHI have been sold. Consequently, its safety and effectiveness are very important to a large number of patients. Raw materials are the source and foundation for production of traditional Chinese medicine injections. In this article, we reviewed the identification of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos, resource distribution, cultivation, quality control, and detection of xenobiotic pollutants, in order to guide the production of high-quality, stable, and pollution-free raw materials. This will be a benefit in ensuring the safety and effectiveness of DHI and reducing the incidence of adverse reactions from the raw materials. By comparing the similarities and differences between the quality standards of Salviae Miltiorrhizae Radix et Rhizoma, Carthami Flos and DHI, we provided some comments for improving the quality standards and post-marketing reevaluation of DHI, and provided some theoretical supports for the production of high-quality herbal raw materials.

Centrin is a member of the EF-hand superfamily that plays critical role in the centrosome duplication and separation. In the present paper, we characterized properties of metal ions binding to Euplotes octocarinatus centrin (EoCen) by fluorescence spectra and circular dichroism (CD) spectra. Changes of fluorescence spectra and α-helix contents of EoCen proved that Tb 3+ and Ca 2+ induced great conformational changes of EoCen resulting in exposing hydrophobic surfaces. At pH 7.4, Ca 2+ (and Tb 3+ ) bond with EoCen at the ratio of 4:1. Equilibrium experiment indicated that Ca 2+ and Tb 3+ exhibited different binding capabilities for C- and N-terminal domains of protein. C-terminal domain bond with Ca 2+ or Tb 3+ ∼ 100-fold more strongly than N-terminal. Aromatic residue-sensitized Tb 3+ energy transfer suggested that site IV bond to Tb 3+ or Ca 2+ more strongly than site III. Based on fluorescence titration curves, we reckoned the conditional binding constants of EoCen site IV quantitatively to be K IV = (1.23 ± 0.51) × 10 8 M −1 and K IV = (6.82 ± 0.33) × 10 5 M −1 with Tb 3+ and Ca 2+ , respectively. Metal ions bond to EoCen in the order of IV > III > II, I.

Glu is highly conserved as the first amino acid of E-helix of the EF-hand protein. In this paper, Glu 101, the first amino acid of E-helix of the third EF-hand motif in Euplotes octocarinatus centrin (EoCen) was mutated to be Lys by the method of site direct mutation. Tb3+ and TNS were used as fluorescence probes in the study of the effect of this mutation to the metal binding characteristic of EoCen by fluorescence spectra. Results indicate that compared with EoCen, the mutation protein (E101K) displays a different Tb3+ binding characteristic and an increased hydrophobic exposure surface. Polyacrylamide gels electrophoresis indicated that the electrophoretic mobilities of EoCen and E101K are distinctly different. It can be deduced that the conformation of EoCen has been altered by this mutation. The general conditional binding constant of Tb3+ to the three loops of EF-hand sites I-III in E101K was calculated to be (5.64+/-0.57)x10(5)M(-1) according to the modified equation of the single binding process.