The scientific investigation on the nature of Chinese materia medica(CMM)has stepped into a critical moment.In this paper,the objectivity of the CMM nature and some other relevant existing problems are discussed based on retrospective and prospective analyses.It could be concluded that there is a duality about the objective connotation of the CMM nature.In order to get a significant breakthrough in near future in the scientific investigation on the CMM nature,the goal,strategy,and methodology should be reconsidered and adjusted.Cold and heat,the most radical and important elements of the CMM nature should be treated as the most principal task in the field.The key scientific issues can be refined to establish a set of objective and scientific evaluating system for the CMM nature,and to elucidate its objective conno- tation.In view of restoration to integration as well as thermodynamic expression,a set of new ideas and methods for the investigation on the CMM nature has been put forward in this paper.

Establishment of bioassay methods is the technical issues to be faced with in the bioassay of Chinese materia medica. Taking the bioassay of Coptis chinensis Franch. as an example, the establishment process and application of the bioassay methods (including bio-potency and bio-activity fingerprint) were explained from the aspects of methodology, principle of selection, experimental design, method confirmation and data analysis. The common technologies were extracted and formed with the above aspects, so as to provide technical support for constructing pattern and method of the quality control for Chinese materia medica based on the dao-di herbs and bioassay.

Biological evaluation is a critical approach by which the quality standardization of traditional Chinese medicines can be linked to the clinical efficacy and safety, which has become an important aspect in the develop-ment of quality standardization of Chinese medicines. In this article, the necessity and feasibility of the biological e-valuation research were analyzed. Further, the main progress achieved as well as the major problems about the re-search and applications of the biological evaluation of traditional Chinese medicines were discussed, to bring forward a point for consideration or discussion.

Objective:To compare the bioactivities of Taohua anti-diarrhea granula prepared by two different methods based on biothermo-kinetics study.Methods:The coarse and fine powder preparations of Taohua decoction were prepared.Microcalorimetry was used to obtain the metabolic curves and biothermo-kinetic parameters of Escherichia coli(E.coli)growth after treated by the two preparations to evaluate their bioactivities.Results:The values of growth rate constant k and maximum heat-out power Pm declined with the concentration increase of the 2 preparations;meanwhile,the values of peak time of the highest peak tm and growth inhibitory ratio I increased with the concentration increase of the two preparations.The results showed that the growth of E.coli was inhibited and the inhibitory effect was strengthened with the concentration increase of the two preparations.The IC50 was 35.9 mg/ml for the coarse powder preparation and 31.4 mg/ml for the fine powder preparation.Aggregate analysis of the metabolic curves and biothermo-kinetic parameters of E.coli indicated that the fine powder preparation of Taohua decoction had stronger inhibitory action against E.coli growth than the traditional coarse powder preparation.Conclusion:Microcalorimetry can be used to examine the bioactivity differences of the fine and coarse preparations of Taohua decoction.Superfine comminution may increase the inhibitory action of Taohua decoction.

The current official quality control approaches meet the challenges from the complexity of herbal medicines.In fact,any herbal medicines containing numerous unknown components,its curative effect usually depends on the whole of herbal medicines,so it is impossible and unnecessary to qualitatively and quantitatively study every component.By investigating the limitations of current quality control approaches for herbal medicines and the difference and similarity in the chemical substantial style as well as quality control pattern of herbal medicines,a new quality control approach for Chinese herbal medicines should be explored and designed.The combination approach of chemical analysis with bioassay is promising to be developed and employed in order to ensure the safety and efficacy of Chinese herbal medicines.

Objective To establish the characteristic biothermokinetic methods in different extract fractions of Radix Isatis using Escherichia coli as model bacteria under biothermodynamics. MethodsMicrocalorimetry was used to obtain the characteristic metabolic growth power-time curves and biothermodynamic parameters of bacteria affected by different extract fractions of Radix Isatis, and evaluate the differences among the different extract fractions of Radix Isatis. Results Different extracting fractions of Radix Isatis could inhibit or promote the growth and metabolism of E. coli in different levels. Water-extract fractions and residues could promote its growth while organic solvent extract fractions did the opposite. Furthermore, with the decline of the solvent polarity, the inhibiting rate of organic solvent extract fraction was decreased and the generation time of bacteria was prolonged. There were steady and distinct differences among the different extract fractions in the characteristic metabolic growth power-time curves, such as the maximum power value's decrease and peak time's delay of water-extract fraction and residue, the specific power enhancement in later stage of organic solvent extract fraction. ConclusionMicrocalorimetry is a new and useful mean for the study on the biothermodynamic parameters and characteristics of Radix Isatis which can be helpful to investigate the other models using microcalorimetry.

To explore the new pattern of Chinese medicine solid preparations (CMSP) in vitro dissolution, a method testing the bio-thermal activity in combination with UPLC was used. Microcalorimetry was used to obtain the characteristic metabolic growth power-time curves and a series of biothermodynamic parameters of the inhibition of Staphylococcus aureus by Yinhuang tablet dissolving solutions at the pH 6.8 (phosphate buffer) dissolution medium at different times. From these results, the cumulative dissolution of Yinhuang tablet based on bio-thermal activity was obtained. The dissolution rates of two components of chlorogenic acid and baicalin were determined by UPLC method. Then f2 similar factor method was used to evaluate the relevance of these two methods. The result showed that f2 values all were more than 50, indicating that there is a good correlation between the two methods of measuring the dissolution rate. It is feasible to determine CMSP in vitro dissolution by using bio-thermal activity, and to provide new evaluation methods for controlling the quality of CMSP.

Objective:To construct an eukaryotic expression vector of enhanced green fluorescence protein(EGFP) gene driven by telomerase catalytic subunit(hTERT) gene promoter and observe the specific expression of EGFP in lung cancer cell lines.Methods:The 1100bp promoter fragment was obtained by enzyme digestion from a recombinant plasmid of pGL3-hTERTp containing the hTERT promoter.The hTERT promoter was then subcloned into the upstream of the report gene EGFP of pEGFP-1 without promoter.The expression vector pEGFP-hTERTp was successfully constructed.The vector pEGFP-N1 containing cytomegalovirus(CMV) promoter was used as a positive control.The vector pEGFP-1 without promoter was used as a negative control.The vectors were transfected into human lung cancer cell lines 95D,NCI-H446,A2,A549,LTEP-a-2,YTMLC and normal MRC-5 through lipofectamine respectively.EGFP expression was detected under the fluorescence microscope.Results:pEGFP-hTERTp was confirmed by enzyme digestion with correct result.That the EGFP expression was detected in all of eight lung cancer cells transfected with pEGFP-hTERTp,but not in MRC cells.By contrast,high intensity EGFP expression was observed in both lung tumor cells and normal cells,which were transfected with pEGFP-N1.Conclusion:The EGFP controlled by hTERT promoter can be expressed specifically in lung cancer cell lines.hTERT promoter may be used as an excellent regulation element in tumor-targeting gene therapy.