Target identification of bioactive compounds is one of the key issues in chemical biology and drug discovery. With the development of science technology, a variety of methods and technologies for target identification have been reported. They can be fundamentally categorized into two approaches: the direct method based on affinity chromatography which is mainly to detect the combination of drug and targets, and the indirect one which is mainly to predict the drug target and action mechanism by physiological reaction and biochemical marker. This review aims to describe the two approaches for target identification.

Objective To investigate the relationship between the content of flavonoids and the key enzyme genes expression in different tissues of Bupleurum chinense and B. scorzonerifolium. Methods The roots, stems, leaves, and fruits of B. chinense and B. scorzonerifolium were used as test materials, determination of flavonoids (rutin, quercetin, kaempferol, and isorhamnetin) in different tissues by HPLC, determination of total flavonoids by UV spectrophotometry, the tissues expression of key enzyme genes (IFS, F3H, and DFR) in flavonoids synthesis was determined by real-time quantitative PCR, correlation analysis was performed with SPSS. Results The content of flavonoids in the aerial parts (stems, leaves, and fruits) of B. chinense and B. scorzonerifolium was significantly higher than that in roots, the content of flavonoids was mainly rutin, and the content of rutin in the leaves of B. chinense leaves was up to 106.961 mg/g; The distribution of total flavonoids in B. chinense and B. scorzonerifolium was obviously different, the content was from high to low: leaves ≥ fruit > stem > root; The expression of B. chinense IFS, F3H, and DFR gene in the aerial parts was much higher than that in roots, IFS gene was significantly positive correlated with rutin (P < 0.05), F3H gene was significantly positive correlated with DFR gene (P < 0.05), but the expression of IFS, F3H, and DFR gene in each tissues of B. scorzonerifolium was at lower level. Conclusion The content of flavonoids in different parts of B. chinense and B. scorzonerifolium was consistent with the expression of flavonoids synthesis key enzyme genes, the differential expression of key enzyme genes regulates the synthesis and accumulation of flavonoids in different tissues.

Objective: To select suitable references genes of Bupleurum scorzonerifolium for tissue expression analyses, and study the tissue expression characteristics of the key enzyme genes of saikosaponins biosynthesis. Methods: Five candidate reference genes including Actin, α-tubulin, β-tubulin, Cyclophilin, and EF-1α were chosen. The stability of these candidate reference genes was investigated by using four softwares (Delta CT, BestKeeper, NormFinder, and GeNorm). The stability of these candidate reference genes was tested and verified by real-time quantitative PCR. Used the stable reference gene, the tissue expression characteristics of the saikosaponins biosynthesis key enzyme genes (HMGR, IPPI, FPS, SS, and β-AS) was analyzed by qRT-PCR. Results: The average expression stability of the five candidate reference genes from high to low was β-tubulin > Cyclophilin > Actin > EF-1α > α-tubulin. Β-tubulin was the most suitable reference gene for tissue expression analysis in B. scorzonerifolium. HMGR expression level was roots > stems and fruits > leaves, IPPI expression level was roots > stems > fruits and leaves, FPS expression level was leaves > roots > stems and fruits, SS expression level was leaves > fruits > roots > stems, β-AS expression level was leaves > roots > fruits > stems. HMGR was significant positive correlated with IPPI, and FPS was significant positive correlated with β-AS (P < 0.05). Conclusion: β-tubulin gene was confirmed as the most suitable reference gene in different tissues of B. scorzonerifolium. It provided a methodological basis for the tissue expression analysis on the functional genes of B. scorzonerifolium. The expression pattern of five key enzyme genes of saikosaponins biosynthesis in different tissues had obvious differentiation, which might be involved in regulating the flow of saikosaponins synthesis and accumulation in various tissues of B. scorzonerifolium.

Objective To explore the effect of ecological factors and the expression of key enzyme genes on the synthesis and accumulation of ginsenosides. Methods Cultivated four-year-old ginseng were used as test materials, the expression of key enzyme genes (HMGR, FPS, SS, SE, DS, β-AS, CYP82D47, CYP716A47) in the biosynthesis of ginsenosides of roots in different growth periods was determined by real-time quantitative PCR, determination of the content of eight ginsenosides (ginsenoside Rg1, Re, Rf, Rb1, Rb2, Rb3, Rc, Rd) in roots by HPLC, the meteorological data were collected by a small weather station, correlation analysis was performed with SPSS. Results The expression of key enzyme genes in the period of flowering to fruit ripening was higher than the root growing after fruit period and the withering period, the expression of key enzyme genes involved in the synthesis of ginsenosides was influenced by each other, and the expression of key enzyme genes in ginseng roots showed a positive correlation with the accumulation of ginsenosides; The contents of ginsenoside Rg1, Rb1, Re, and Rc were higher in the roots of ginseng, eight kinds of monomer ginsenosides content dynamic changes trend is different; Temperature, photosynthetically active radiation, soil water potential are important ecological factors for ginsenosides synthesis in roots, temperature was significantly negatively correlated with ginsenoside Rb1 and Rd (P < 0.05), PAR can significantly promote the formation of ginsenoside Rg1 (P < 0.05), soil water potential was significantly negatively correlated with ginsenoside Rb1 (P < 0.05); Grey correlation analysis results showed that the major ecological factors that influenced ginsenosides content in ginseng roots were temperature, PAR and relative humidity, the grey correlation between the expression of the key enzyme genes with the content of ginsenosides is less than ecological factors with the content of ginsenosides, under the guidance of ecological factors, the expression of the key enzyme genes regulate the synthesis and accumulation of ginsenosides. Conclusion The dynamic changes of the expression of key enzyme genes and the content of ginsenosides in ginseng were determined, it provides a theoretical basis for elucidating the physiological and ecological mechanism of ginsenoside synthesis and the quality control of Radix Ginseng.

Objective: The research indicated that the nature of Chinese medicine is mainly related to body's substance and energy metabolism. The purpose of the study is to elucidate the substance basis for warm nature of Poria cocos (called Fuling (FL) in Chinese). Methods: In terms of the effects of its separated fractions on the substance and energy metabolism in rat models of cold-deficiency with Aconiti Lateralis Radix Praeparata (called Fuzi (FZ) in Chinese), with hot nature, as reference drug. Biochemical indexes in the material metabolism, energy metabolism, endocrine system, nervous system and nucleotide system were determined, then analyzed by additive, cluster and principal component analysis (PCA). Results: The medicinal natures of oligosaccharides and amino acids fractions were attributable to plain and crude polysaccharides, volatile oils and triterpenoids fractions were attributable to mild warm. Conclusion: The nature of FL was regarded as mild warm based on the old records of Chinese medicine and fractions of crude polysaccharides, volatile oils and triterpenoids might be the main substance basis for the warm nature of FL. It is the first time that substance basis of FL was elucidated from view point of medicinal nature.

Cistanches Herba, known as "Ginseng of the desert", is authenticated from the dried succulent stems of Cistanche deserticola and Cistanche tubulosa. As a famous remedy in China for tonic the kidney, it is used to treat "kidney-deficiency syndrome"-induced diseases such as infertility, forgetfulness, hearing lost, chronic constipation, etc.. As various biological activities, including anti-aging, antioxidant, estrogenic, anti-osteoporotic, and anti-inflammation effects, have been discovered, here we reviewed Cistanches Herba in biological characteristics, chemical constituents, and pharmacological activities.

Objective To establish an HPLC method for simultaneous determination of loganin, chiratin, secoxyloganin, rosmarinic acid and calceolarioside B in Shuangxiangpaishi granules. Method HPLC colunmn was Alltima C18 column (4.6 mm×250 mm, 5 μm); mobile phase consisted of acetonitrile-methanol (1: 3) (A) -0.1% phosphate acid solution (B) with gradient elution; the column temperature was 35°C; the flow rate was 1.0 ml/min; all the injection volume was 20 μl; loganin, chiratin and secoxyloganin were detected at 240 nm, rosmarinic acid and calceolarioside B were detected at 330nm. Results The above mentioned five main ingredients had linearity in the given concentration range at 5.91-118.20 μg/ml (r=0.9995), 4.10-82.00 μg/ml (r=0.9991), 8.13-162.60 μg/ml (r=0.9993), 12.57-251.40 μg/ml (r=0.9998), 4.95-99.00μg/ml (r=0.9997), respectively. The average recoveries (n=6) and RSD were 96.88 (1.31), 99.09 (1.47%), 98.29 (1.59), 98.82 (1.42) and 97.51 (0.86), respectively. Conclusion This dual-wavelength HPLC method could simultaneously determine the contents of the five ingredients in Shuangxiangpaishi granules and can be used to evaluate their quality. The method is simple, accurate, sensitive and repeatable. It could be used as an efficient strategy for systematic quality evaluation of Shuangxiangpaishi granules.

OBJECTIVE: To review the chemical constituents from Camptotheca acuminate Decne and report bioactivities, which provides detailed information for further investigations and applications of C. acuminate, a plant resource endemic to China. METHODS: Based on 39 domestic and foreign literatures, the analysis, induction and summarization of chemical constituents and bioactivities of C.acuminate. RESULTS and CONCLUSION: The major chemical components of C. acuminate are alkaloids, ellagic acid, and flavonoids. The crude extract of C. acuminate shows anti-tumor, immunosuppression and anti-virus activities.

OBJECTIVE: To establish the HPLC fingerprint of traditional Chinese medicine Huangkui Capsules, thus to provide an approach and a basis for the quality control in production. METHODS: The analytical column was Thermo scientific Hypersil GOLD. The mobile phase was composed of acetonitrile (A) and 0.2% phosphoric acid (B) with gradient elution. The flow rate was 0.8 mL · min-1, and the detection wavelength was set at 360 nm with the column temperature maintained at 30°C. The fingerprints of the extracts from the raw materials, intermediate product, and Huangkui Capsules were compared. RESULTS: The similarities of the chromatograms of 10 different lots of Huangkui Capsules were over 0.95. The correlation was good between the raw materials, intermediate product and Huangkui capsules. CONCLUSION: This validated method is suitable for the quality evaluation and quality control of Huangkui Capsules.

To explore the medication rule of Chinese materia medica (CMM) in the treatment of pulmonary fibrosis, and provide reference for the prevention and treatment of pulmonary fibrosis in the novel coronavirus pneumonia. Using Excel 2013 and SPSS Statistics 23.0, the literatures with the theme of “pulmonary fibrosis” that met the screening criteria were searched, and the association rule analysis was carried out. Frequency analysis showed that the occurrence frequency of Astragali Radix (148, 45.82%), Glycyrrhizae Radix et Rhizoma (124, 38.39%) and Salviae Miltiorrhizae Radix et Rhizoma (102, 31.58%) was the highest in the clinical treatment literature. In the statistics of nature and flavor, warm (10 times, 50.00%) and neutral (six times, 30.00%) were the most common. The highest frequency of meridian tropism was found in the spleen (11 times, 25.58%), lung (10 times, 23.26%) and sweet (11 times, 36.67%). In the analysis of association rules, six groups of TCM combinations with high frequency correlation strength were obtained. The high frequency and high correlation CMM combinations obtained from this data analysis were mainly effective in strengthening the spleen and invigorating qi, and warming the spleen and lung, which can be used to prevent the early patients from developing to the critical stage and prevent the patients from reinfection due to low immunity in the convalescence stage.