The relationship between the drug effect and the time interval from operation or onset of hepatitis or hepatic damage to the beginning of the administration of Chinese herb medicines allowed us to postulate the usual clinical course of the disease as follows from the viewpoint of traditional Chinese medicine: namely two months after surgery or onset of the disease being the Yang stage, the four months after the Yang stage being the Yang-Yin transitional stage and after this being the Yin stage. The Yin stage was further divided into earlier and later periods, the border line being five years after the operation or onset of the disease.
In the Yang stage the drug effect was mostly excellent, and the medicines used were mainly for Qi-fen and Re-zheng, as well as the medicines used in the Yang-Yin transitional stage when healing of the excellent cases and increase of the cases with good effect were characteristic. In the earlier Yin stage the cases with no effect began to increase, and the medicines used ranged from Qi-fen to Ying-fen . In the later Yin stage the cases with excellent effect disappeared, and the cases with good or no effect were dominant. In this stage the medicines for Xue-fen were sometimes added to the medicines for Qi-fen or Ying-fen, and relative increase of the medicines for Xu-han-zheng was noticed.

Objective: To investigate the effect of the roselle calyx extract (RCE) (Hibiscus sabdariffa L.) on the in vitro viability and biofilm formation ability of oral pathogenic bacteria. Methods: RCE was prepared by soaking roselle calyx powder with ethyl alcohol for 24 h at room temperature. After centrifugation, the extract was lyophilized. Then, the extract was dissolved in phosphate-buffered saline, the pH was adjusted, and the extract was aseptically filtered. We used Streptococcus mutans, Streptococcus sanguinis, Lactobacillus casei, Actinomyces naeslundii, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis and Prevotella intermedia in this study. The antibacterial activity of the RCE was determined by treating the cells of these bacteria with the extract for 10 or 20 min at room temperature. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration was determined using the microdilution method, and the effect of the RCE on the ability to form biofilm was determined using a polystyrene micro plate assay. In addition, we used the WST-1 assay to determine the cytotoxicity of the RCE on HGF, Ca9-22 and KB cells. Results: The RCE had antibacterial activity against oral bacteria used in this study. In particular, most significant antibacterial activity was observed against Fusobacterium nucleatum, Prevotella intermedia and Porphyromonas gingivalis. The MIC and minimum bactericidal concentration were 7.2 mg/mL-28.8 mg/mL and 14.4 to >57.6 mg/mL. The RCE had an inhibitory effect on biofilm formation at the MIC and sub-MIC levels. In addition, the RCE had low cytotoxic effects on HGF, Ca9-22 and KB cells. Conclusions: Thus, our results indicate that the RCE may be used for preventing oral diseases.