[Mechanisms of Fuke Qianjin Capsules in treatment of pelvic inflammatory disease based on GC-MS metabolomics].

Pelvic inflammatory disease( PID) rat model was induced by the mixture of Escherichia coli,Staphylococcus aureus,and Streptococcus hemolytic-ß． Gas chromatography-mass spectrometry( GC-MS) based metabolic profiling method was combined with multivariate statistical analysis,such as PCA,PLS-DA and OPLS-DA to analyze endogenous small molecule metabolites in serum of rats after treatment of Fuke Qianjin Capsules. The results showed that Fuke Qianjin Capsules could significantly improve the inflammatory pathological characteristics and tissue damages in model rats. Based on the principle of VIP>1 and P<0. 05,a total of 6 different metabolic biomarkers were identified,including L-valine,L-isoleucine,L-threonine,butanedioic acid,serine and D-glucose,respectively.The contents of these six different metabolites were significantly reversed after administration. Further analysis of the metabolite pathways through KEGG database showed that Fuke Qianjin Capsules achieved the effect possibly through glycine,serine and threonine metabolism,aminoacyl-tRNA biosynthesis and valine,leucine and isoleucine biosynthesis. Therefore,this study came to the conclusion that Fuke Qianjin Capsules can be used in the treatment of mixed bacteria induced pelvic inflammatory disease possibly by regulating amino acid and its derivative metabolism.

[Analysis on multiple pharmaceutical ingredients and antioxidant capacities of Prunellae Spica based on multivariate statistical analysis].

Prunellae Spica is a perennial edible and medicinal plant, rich in antioxidant substances. Total flavonoids (TFC), Phenolics (TPC), triterpenoids (TSC), polysaccharides (PC) and their antioxidant capacities (by the FRAP, DPPH and ABTS⁺ methods) of ethyl acetate fraction, n-butanol fraction and other fractions of aqueous extract from Prunellae Spica were investigated in this study. Then the multivariate statistical method was adopted to analyze the relationship between the multiple pharmaceutical ingredients and antioxidant capacities of Prunellae Spica. The results showed that ethyl acetate fraction had relatively high concentration of TFC (0.61±0.10) g·g⁻¹DW, TPC (0.52±0.09) g·g⁻¹DW, and TSC (0.21±0.03) g·g⁻¹DW, with high scavenging capacity of DPPH (3.1±0.38) mmol·L⁻¹·g⁻¹DW and FRAP (2.56±0.35) mmol·L⁻¹·g⁻¹DW. Hierarchical clustering analysis (HCA) and principal component analysis (PCA) results indicated the information from chemical compositions and antioxidant capacity can represent the "differences" of different fractions. Canonical correlation analysis (CCorA) revealed a high positive correlation between the amounts of multiple chemical compositions and the antioxidant capacities (r=0.970 0), and the first canonical variate had been reached. Moreover, ABTS⁺ method showed a low response to the compositions of different fractions, so this method may not be suitable for evaluation of Prunellae Spica antioxidant capacities, while DPPH evaluation method was more suitable for TSC and TPC. The results of this study have important reference significance for the evaluation method on antioxidant activity of Prunellae Spica in the field of food or medicine as well as for the development of related extracts.

[Extraction of flavonoids in Prunella vulgaris based on deep eutectic solvent method：application of new green solvent].

Flavonoids have attracted much attention due to their good anti-inflammatory, anti-oxidation and anti-tumor effects. At present, the extraction of flavonoids is mainly based on organic solvent, while the researches on the use of green and safe solvents are quite limited. Therefore, in the present study, different types of deep eutectic solvents (DESs) were applied to investigate their effect on extraction of flavonoids and optimize the process, also investigate the recovery efficiency of DESs and evaluate the recovery method for total flavonoids. The extraction yield of the total flavonoids acted as the comprehensive evaluation indexes, and a central composite design (CCD) of response surface methodology (RSM) was employed to further optimize the alcohol-based DES extraction conditions. The results showed that the optimized extraction conditions were as follows: water-DES ratio of 27%, solid-liquid ratio of 15 mL·g⁻¹, extraction temperature of 83 °C and extraction time of 42 min in ChCl-glycerol at 1:4 ratio. Under these conditions, the mean experimental value of the extraction yield (75.05 mg·g⁻¹) corresponded well with the predicted value (77.86 mg·g⁻¹). Moreover, these experimental results showed more advantages such as in higher efficiency, economy and environmental protection as compared with previously reported conventional extraction methods. In addition，the recovery yield of the total flavonoids from the DESs extraction solution achieved 97.88% by using AB-8 macroporous resin, and 88.12% desorption ratio can be achieved by 100% ethanol with 5 times resin content. After the above treated DESs were collected, the extraction yield with the same method reached 95.23%, indicating that the method of macroporous resin can be used for efficient and simple recovery and reuse. This study suggests that DESs can be used as a kind of sustainable and efficient natural extraction solvents for extraction of flavonoids from Prunella vulgaris.

[Application of random forest algorithm in fingerprint of Chinese medicine: different brands of Xiasangju granules as example].

To establish a random forest algorithm for identifying and classifying different brands of Xiasangju granules, and provide effective reference for identifying multi-index complex fingerprint. HPLC method was used to collect the fingerprint of 83 batches of Xiasangju granules from different manufacturers. The classification of Xiasangju granules samples based on chromatographic fingerprints was identified by chemometric methods including principal component analysis (PCA), partial least squares discriminate analysis (PLS-DA) and random forest analysis (RF). The superiority of the above three chemometric methods was compared. The results showed that the fingerprints of 83 batches of Xiasangju granules were established in this study. PCA could only explicate 56.52% variance contribution rate and could not completely classify the samples; PLS-DA analysis was superior to PCA, explicating 63.43% variance contribution rate and could obtain certain separation; RF could well classify the samples into 3 types, and the predication accuracy of the proposed method was 96.5%. Therefore, The results indicate that RF combined with HPLC fingerprint could effectively construct traditional Chinese medicine quality control and analysis system.

Enzymatic Extraction, Purification, and Characterization of Polysaccharides from Penthorum chinense Pursh: Natural Antioxidant and Anti-Inflammatory.

Penthorum chinense Pursh (PCP) is a kind of functional food or medicine for liver protection. In the present work, Plackett-Burman design, steepest ascent method, and response surface methodology (RSM) were employed to obtain maximum total sugar yield. The experimental yield of 6.91% indicated a close agreement with the predicted yield of 7.00% of the model under optimized conditions. The major polysaccharide fraction (PCPP-1a) from PCPP was purified and identified as acidic polysaccharides with a high content of uronic acid (FT-IR, UV, HPGPC). PCPP had similar monosaccharide profile with PCPP-1a but was rich in galacturonic acid (HPLC). Both of PCPP and PCPP-1a possessed strong hydroxyl radical scavenging, DPPH radical scavenging, and Fe2+ chelating activities. Moreover, they were revealed to show strong anti-inflammatory activities by inhibiting NO, TNF-α, and IL-1ß release compared to LPS treatment in RAW264.7 cells. These data suggest that the polysaccharides from PCP could be potential natural products for treating ROS and inflammatory-related diseases.

Determination of Total Flavonoids Contents and Antioxidant Activity of Ginkgo biloba Leaf by Near-Infrared Reflectance Method.

BACKGROUND: Total flavonoids content (TFC) is one of the most important quality indexes of Ginkgo biloba leaf, and it is concerned with total antioxidant activity. Near-infrared spectroscopy (NIR) method has showed its advantages in fast, accurate, qualitative, and quantitative analysis of various components in many quality control researches. In this study, a calibration model was built by partial least squares regression (PLSR) coupling with NIR spectrum to quantitatively analyze the TFC and total antioxidant activity of Ginkgo biloba leaf. RESULTS: During the model establishing, some spectrum pretreatment and outlier diagnosis methods were optimized to establish the final model. The coefficients of determination (R2 ) for TFC and total antioxidant activity prediction were 0.8863 and 0.8486, respectively; and the root mean square errors of prediction (RMSEP) were 2.203 mg/g and 0.2211 mM/g, respectively. CONCLUSION: These results showed that NIR method combined with chemometrics is suitable for quantitative analysis of main components and their activities and might be applied to quality control of relevant products.

Extraction, Characterization, Antioxidant, and Immunostimulatory Activities of Polysaccharides from Hedyotis corymbosa.

In this study, optimization of enzyme-assisted extraction, purification, characterization, and its bioactivities of polysaccharides from Hedyotis corymbosa (HCP) was investigated. It was found that the optimum extraction conditions were 3% of enzyme concentration (X 1 ), 30 of liquid-to-solid ratio (X 2 ), 56°C of extraction temperature (X 3 ), 200W of ultrasonic power (X 4 ), 10 min of extraction time (X 5 ), and 5 of pH value (X 6 ). Under optimum conditions, the experimental yield (4.10 ± 0.16%) was closed to the predicted value (4.02%). The crude HCP was further purified using DEAE-52 and Sephadex G-150 gel column, and a major polysaccharide fraction from HCP, designed as HCP-1a with molecular weight of 33.9 kDa, was obtained. The HCP and HCP-1a were characterized by chemical analysis, FT-IR, and HPLC. For antioxidant activities in vitro, HCP possessed strong hydroxyl radical scavenging, DPPH radical scavenging, and Fe2+ chelating activities. In subsequent immunostimulatory studies, significantly decreased NO, IL-1ß, and TNF-α concentrations were observed in both of HCP and HCP-1a treated RAW264.7 cells. Therefore, this study may indicate some insights into the application of polysaccharides from Hedyotis corymbosa as potential natural antioxidants and immunostimulants.