Selecting and Characterizing Tyrosinase Inhibitors from Atractylodis macrocephalae Rhizoma Based on Spectrum-Activity Relationship and Molecular Docking.

Atractylodis macrocephalae Rhizoma (AMR) is a famous classical Chinese traditional medicine (CTM), which has been used as a tonic for many diseases for thousands of years. In ancient China, it was used as a supplementary food for beauty in the palace. In preliminary studies, the function of whitening skin and the significant inhibiting effect on tyrosinase (TYR) which is the reactive enzyme in the composition of melanin of AMR were discovered, and the relevant research was rarely reported. In this study, high-performance liquid chromatography (HPLC) along with partial least squares regression analysis (PLS) was applied to survey the coherence between the chemical constituents and the inhibiting activity of 11 batches of AMR on TYR activity. The results of PLS showed that the chromatographic peaks 11 (atractylenolide III) and 15 could be important effective ingredients of the inhibition TYR activity as ascertained by spectrum-activity relationships. Furthermore, TYR inhibitory activity of atractylenolide III was validated by in vitro test by ß-arbutin served as a positive control drug. The results of the in vitro test and the molecular docking showed that atractylenolide III has high TYR inhibitory activity and could link to the residues in TYR catalytic pocket. Therefore, bioassay, molecular docking, and spectrum-activity relationships are appropriate for linking the quality of samples with pharmaceutical-related active ingredients. And our studying would lay a theoretical foundation for applying the water extracts of AMR in whitening cosmetics.

[Determination and analyse of soil fertility of Pseudostellariae Radix planting base of Shibing County and Huangping County in Guizhou Province].

OBJECTIVE: To determine soil fertility of the Pseudostellariac Radix planting base of Shibing County and Huangping County in Guizhou Province, and to provide experimental basis for soil improvement and balanced fertilization. METHODS: 12 soil samples from Shibing County and 10 soil samples from Huangping County were involved in the detection. RESULTS: In the soil samples from Shibing County, the contents of organic matter, total N, available N, total P, available P and available K were higher, while the content of total P was at middle level. In the soil samples from Huangping County, the contents of total N, available P, total K and available K were higher,the contents of organic matter, total K and available N were at middle level. CONCLUSION: The level of soil fertility of the Pseudostellariae Radix planting base in Guizhou Province is common. The soil fertility of Shibing County is superior to that of Huangping County. The soil pH value of Shibing and Huangping County is low. In Shibing County, lime, farmyard manure and neutral fertilizer can be used to increase the pH of soil. Potash fertilizer can be increased, while nitrogen fertilizer and phosphate fertilizer can be reduced in the soil of Shibing County. Potash fertilizer and phosphate fertilizer will be increased, while nitrogen fertilizer can be reduced in the soil of Huangping County. The soil fertility can be improved by these measures.

[Study on HPLC fingerprint of miao medicine Ardisia japonica].

OBJECTIVE: To establish the HPLC fingerprint of Ardisia japonica. METHODS: HPLC analysis was performed on a Diamonsil C18 column (250 mm x 4.6 mm, 5 microm) with the eluting system of gradient consisted of methanol-0.1% H3PO4. The flow rate was 1.0 mL/min. The column temperature was maintained at 35 degrees C and the detection wavelength was 250 nm. RESULTS: The HPLC fingerprint was established and 10 batches of samples with 32 common peaks were compared. Four peaks were identified as gallic acid, bergenin, chlorogenic acid and quercitrin. CONCLUSION: The method with good reproducibility is simple and accurate, which can be used for determination of HPLC fingerprint and quality control of Ardisia japonica. It provides scientific basis of future study of Ardisia japonica.

[Study on immunological effect of housefly protein in mice].

OBJECTIVE: To study the immunological effect of the housefly protein in mice. METHODS: The housefly protein was given orally in normal and Cyclophosphamide (CY) immunosuppressive mice, and then the indexes of the clearance rate of carbon particles and the serum hemolysin against SRBC were detected. RESULTS: The housefly protein could increase HC50 significantly in the normal mice, but could not in the CY immunosppressive mice. It could increase phagocytic index significantly in the normal mice, but could not in the CY immunosuppressive mice. CONCLUSION: The results suggest that the housefly protein can promote humoral immunity functions of normal mice. It has distinct preventive and cure effect on the CY immunosuppressive.

[Isolation and purification of antibacterial peptides from the larvae secretion of housefly and the characteristics].

OBJECTIVE: To isolate and purify the antibacterial peptides from larvae secretion of housefly (Musca domestica) and study their partial characteristics. METHODS: Protein isolation and purification were performed by routine process, namely, ultrafiltration, solid phase extraction (SPE) and reversed-phase high-performance liquid chromatography (RP-HPLC). The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the antibacterial peptides were examined. The antibacterial effect of peptides was studied in nutritive medium with different pH value(pH 5.0-10.0), divalent cations (Mg2+: (0.5 x 10(-3) - 10.0 x 10(-3))mol/L, Na+, K+: (10 x 10(-3) - 100 x 10(-3))mol/L), and serum content (12.5%-75%). RESULTS: Molecular weight of the peptides was about Mr 3 000-30 000 after ultrafiltration. The fractions eluted with 20%, 30%, 70%, and 80% of acetonitrile (ACN) all showed antibacterial activity by solid phase extraction. The fractions eluted with 70% ACN showed strongest and stable st antibacterial activity which was further purified by RP-HPLC. Two sub-fractions appeared at around RT 15.5 min and 18.5 min were obtained with antibacterial activity. The MIC to those standard Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis was 32.7380, 16.3688, 65.4750 and 32.7380 microg/ml respectively. In the nutritive medium of pH 6.0-9.0, different divalent cations and serum content, the increment of A570 in experiment groups was less than 0.05, while that of the control group was greater than 0.3 (P<0.01). CONCLUSION: SPE and RP-HPLC have been effective in purifying the antibacterial peptides which show strong activity in different conditions.