[Content of secondary metabolites and antioxidant activities of Lonicera japonica flowers at different developmental stages from Shandong province].

This study established an ultrasound-assisted extraction-high performance liquid chromatography method for simulta-neously determinining the content of 11 bioactive compounds including iridoids, phenolic acids, and flavonoids in Lonicera japonica flowers. The flowers at six stages from the rice bud stage(ML) to the golden flower stage(JH) of L. japonica varieties 'Sijuhua' and 'Beihua No.1' in two planting bases in Shandong province were collected. The established method was employed to determine the content of 11 target compounds, on the basis of which the dynamics of active components in L. japonica sampels during different development stages was investigated. The correlation analysis was carried out to reveal the correlations of the content of iridoids, phenolic acids, and flavonoids. Furthermore, the antioxidant activities of samples at different developmental stages were determined, and the relationship between antioxidant activity and chemical components was analyzed by the correlation analysis. The results showed that the total content of the 11 components in 'Sijihua' changed in a "W" pattern from the ML to JH, being the highest at the ML and the second at the slight white stage(EB). The total content of 11 compounds in 'Beihua No.1' was the highest at the ML and decreased gra-dually from the ML to JH. The samples of 'Sijihua' had higher content of iridoids and lower content of phenolic acids than those of 'Beihua No.1'. The content of flavonoids and phenolic acids showed a positive correlation(R~2=0.90, P<0.05) in 'Sijihua' but no obvious correlation in 'Beihua No.1'. The antioxidant activity and phenolic acid content showed positive correlations, with the determination coefficients(R~2) of 0.84(P<0.05) in 'Beihua No.1' and 0.73(P<0.05) in 'Sijihua'. The antioxidant activity of both varieties was the strongest at the ML and the second at the EB. This study revealed that the content dynamics of iridoids, phenolic acids, and flavonoids in 'Sijihua' and 'Beihua No.1' cultivated in Shandong province during different developmental stages. The results indicated that the antioxidant activity of L. japonica flowers was significantly correlated with the content of phenolic acids at different deve-lopmental stages, which provided a basis for determining the optimum harvest time of L. japonica flowers.

Subcritical water extraction of polysaccharides from Gastrodiae Rhizoma: optimization, characterization and in vitro hepatoprotective activity.

Subcritical water extraction (SWE) is an efficient and eco-friendly technology that rapidly extracts valuable compounds from natural materials. In this study, response surface methodology (RSM) was utilized to determine the optimal extraction conditions for Gastrodiae Rhizoma using SWE (GRP-S). The optimum conditions were found to be 161 °C extraction temperature, 41 min extraction time, and a solid-to-liquid ratio of 1.55 mg/mL. Under these optimal conditions, the experimental yield of GRP-S was 66.32% ± 0.10% (n = 3), demonstrating a significant increase compared to hot water reflux extraction (HWE) in the extraction yield of polysaccharides. Characterization studies employing SEM, FT-IR, and HPAEC-PAD confirmed the differences between GRP-S and GRP-H (GRP obtained by HWE). Furthermore, both GRP-S and GRP-H exhibited a significant ability to protect HepG2 cells from ethanol-induced damage, with GRP-S showcasing a superior effect. The widespread adoption of SWE technology can lead to high GRP content in extracts and promote the green and sustainable development of natural products extraction processes.

[Application of bioassay in quality evaluation of Chinese medicine:a review].

The quality of Chinese medicine is the foundation of the clinical effects and industrial development. Component analysis ensures the consistency and stability of medicinals, but fails to evaluate the clinical efficacy. Bioassay is an analytical method to evaluate the effect of a substance on living organisms, tissues, or cells, which is an optimal option for assessing the quality of Chinese medicine. Bioassay of Chinese medicine starts early but progresses slowly. At the moment, it has attracted the interest of scholars. However, no systematic research is available. This study aims to summarize the research on the application of bioassay in quality evaluation of Chinese medicine, focusing on the application of key techniques and experimental systems in bioassay in heat-clearing and blood-activating and stasis-eliminating Chinese medicine and the common problems. Meanwhile, suggestions were proposed in terms of the association with clinical efficacy and chemical analysis and the status quo of biological assay. This study is expected to promote the study and application of bioassay.

Eight-gene metabolic signature related with tumor-associated macrophages predicting overall survival for hepatocellular carcinoma.

BACKGROUND: In recent years, the relationship between tumor-associated macrophages (TAMs) and solid tumors has become a research hotspot. This study aims to explore the close relationship of TAMs with metabolic reprogramming genes in hepatocellular carcinoma (HCC) to provide new methods of treatment for HCC. METHODS: The study selected 343 HCC patients with complete survival information (survival time > = 1 month) in the Cancer Genome Atlas (TCGA) as study subjects. Kaplan-Meier survival analysis assisted in determining the relationship between macrophage infiltration and overall survival (OS), and Pearson correlation tests were used to identify metabolic reprogramming genes (MRGs) associated with tumor macrophage abundance. Lasso regression algorithms were used on prognosis-related MRGs identified by Kaplan-Meier survival analysis and univariate Cox regression analysis to construct a risk score; another independent cohort (including 228 HCC patients) from the International Cancer Genome Consortium (ICGC) was used to verify prognostic signature externally. RESULTS: A risk score composed of 8 metabolic genes could accurately predict the OS of a training cohort (TCGA) and a testing cohort (ICGC). The risk score could be widely used for people with different clinical characteristics, and it is a predictor that is independent of other clinical factors that affect prognosis. As expected, compared with the low-risk group, the high-risk group exhibited an obviously higher macrophage abundance, together with a positive correlation between the risk score and the expression levels of three commonly used immune checkpoints (PD1, PDL1, and CTLA4). CONCLUSION: Our study constructed and validated a novel eight-gene signature for predicting HCC patient OS, which may contribute to clinical treatment decisions.

Purification of hypocrellins from Shiraia bambusicola by coordinated high-speed countercurrent chromatography using cupric chloride as a complexing agent.

Hypocrellins are anthraquinone that can act as excellent photosensitizers for photodynamic therapy. In the present work, we found that high-speed countercurrent chromatography using cupric chloride as a complexing agent effectively separated hypocrellins from Shiraia bambusicola extract. The optimal two-phase solvent system consisted of petroleum ether/ethyl acetate/methanol/water (7:3:5.5:4.5, v/v/v/v), with 0.01 mol/L cupric chloride in the lower phase at pH of 2.45. This lower phase served as the mobile phase, whereas the upper phase acted as the stationary phase. Employing a continuous separation mode, three continuous injections were found to allow the purification of 1.2 g of crude extract in approximately 12 h. Hypocrellin B (10.8 mg), hypocrellin A (16.2 mg), and hypocrellin C (15.6 mg) were obtained from this process. Simulation of complexation of hypocrellin A with divalent copper ion by computational chemistry calculations indicated that three pairs of hydroxyl and carbonyl groups in hypocrellin A had similar binding energies, and demonstrated that hypocrellin A and B owned different metal-to-ligand ratios as compared to hypocrellin C. These factors could modify the partitioning of these compounds in two-phase solvent system, and resulting in a suitable separation factor. This method would also be used to purify other anthraquinones from natural products.

Effect of inorganic salt on partition of high-polarity parishins in two-phase solvent systems and separation by high-speed counter-current chromatography from Gastrodia elata Blume.

Parishins are high-polarity and major bioactive constituents in Gastrodia elata Blume. In this study, the effect of several inorganic salts on the partition of parishins in two-phase solvent systems was investigated. Adding ammonium sulfate, which has a higher solubility in water, was found to significantly promote the partition of parishins in the upper organic polar solvents. Based on the results, a two-phase solvent system composed of butyl alcohol/acetonitrile/near-saturated ammonium sulfate solution/water (1.5:0.5:1.2:1, v/v/v/v) was used for the purification of parishins by high-speed counter-current chromatography. Fractions obtained from high-speed counter-current chromatography were subjected to semi-preparative high-performance liquid chromatography to remove salt and impurities. As a result, parishin E (6.0 mg), parishin B (7.8 mg), parishin C (3.2 mg), gastrodin (15.3 mg), and parishin A (7.3 mg) were isolated from water extract of Gastrodia elata Blume (400 mg). These results demonstrated that adding inorganic salt that has high solubility in water to the two-phase solvent system in high-speed counter-current chromatography was a suitable approach for the purification of high-polarity compounds.

Ultra-high-pressure-assisted extraction of wedelolactone and isodemethylwedelolactone from Ecliptae Herba and purification by high-speed counter-current chromatography.

Ultra-high-pressure extraction combined with high-speed counter-current chromatography was employed to extract and purify wedelolactone and isodemethylwedelolactone from Ecliptae Herba. The operating conditions of ultra-high-pressure extraction were optimized using an orthogonal experimental design. The optimal conditions were 80% aqueous methanol solvent, 200 MPa pressure, 3 min extraction time and 1:20 (g/mL) solid-liquid ratio for extraction of wedelolactone and isodemethylwedelolactone. After extraction by ultra-high pressure, the extraction solution was concentrated and subsequently extracted with ethyl acetate; a total of 2.1 g of crude sample was obtained from 100 g of Ecliptae Herba. A two-phase solvent system composed of petroleum ether-ethyl acetate-methanol-water (3:7:5:5, v/v) was used for high-speed counter-current chromatography separation, by which 23.5 mg wedelolactone, 6.8 mg isodemethylwedelolactone and 5.5 mg luteolin with purities >95% were purified from 300 mg crude sample in a one-step separation. This research demonstrated that ultra-high-pressure extraction combined with high-speed counter-current chromatography was an efficient technique for the extraction and purification of coumestans from plant material.

Evaluation of a Nondestructive NMR and MRI Method for Monitoring the Drying Process of Gastrodia elata Blume.

Gastrodia elata Blume (G. elata) is a prominent traditional herb and its dry tuber is officially listed in the Chinese Pharmacopoeia. To ensure the quality of dried G. elata, the establishment of a nondestructive and convenient method to monitor the drying process is necessary. In this study, a nondestructive low-field nuclear magnetic resonance (LF-NMR) and magnetic resonance imaging (MRI) method was introduced to monitor the drying process of G. elata. Three water states (bound, immobilized, and free) in G. elata samples were investigated through multiexponential fitting and inversion of the NMR data. The variation and distribution of the three water states during drying were monitored by LF-NMR, and the spatial distribution of water and internal structural changes were analyzed by MRI. Linear analysis of the moisture content, L* (lightness), b* (yellowness), and NMR parameters showed good correlations among them. Furthermore, partial least squares regression (PLSR) model analysis, which takes into account all NMR parameters, also showed good correlations among these parameters. All results showed that LF-NMR was feasible and convenient for monitoring moisture content. Therefore, LF-NMR and MRI could be used to monitor the moisture content nondestructively in the drying process of Chinese traditional herbs.

Preparative Separation of Monoterpenoid Indole Alkaloid Epimers from Ervatamia yunnanensis Tsiang by pH-Zone-Refining Counter-Current Chromatography Combined with Preparative High-Performance Liquid Chromatography.

An effective method was developed for the preparative separation and purification of monoterpenoid indole alkaloid epimers from Ervatamia yunnanensis Tsiang using a combination of pH-zone-refining counter-current chromatography and preparative high-performance liquid chromatography. With this method, two pairs of MIA epimers including ervatamine (72 mg, 1), 20-epi-ervatamine (27 mg, 4), dregamine (95 mg, 2), tabernaemontanine (129 mg, 3), along with two MIAs, apparicine (112 mg, 5) and isovoacangine (15 mg, 6), were successfully purified from 2.1 g crude extract of E. yunnanensis, each with a purity of over 95% as determined by HPLC. The structures of the MIAs were identified by ESI-MS, 1D, and 2D NMR.

Chemical Constituents from Scindapsus officinalis (Roxb.) Schott. and Their Anti⁻Inflammatory Activities.

One new monoterpene glycoside (1), one new phenyl glycoside (2), one new caffeoyl derivative (3), were isolated from Scindapsus officinalis (Roxb.) Schott., along with four known compounds (4⁻7). Structures of the isolated compounds were elucidated by extensive analysis of spectroscopic data, especially 2D NMR data and comparison with literatures. All isolates were evaluated for anti-inflammatory activity against nitric oxide (NO) production in vitro. Compounds 3 and 7 exhibited moderate inhibitory effects on NO production with IC50 values of 12.2 ± 0.8 and 18.9 ± 0.3 µM, respectively.

Separation of Seven Polyphenols from the Rhizome of Smilax glabra by Offline Two Dimension Recycling HSCCC with Extrusion Mode.

An offline two-dimensional recycling high-speed countercurrent chromatography (2D R-HSCCC) strategy with extrusion mode was developed for isolating polyphenols from the rhizome of Smilax glabra. Firstly, the ethyl acetate extract was divided into two fractions, Fr.1 and Fr.2, by silica gel column chromatography. Then, HSCCC was applied to separate polyphenols from the two fractions using a solvent system consisting of petroleum ether-ethyl acetate-methanol-water (1:3:0.5:5, v/v). Fifty milligrams of Fr.1 was separated by conventional HSCCC, yielding 5-O-caffeoylshikimic acid (1, 15.8 mg) and taxifolin (2, 4.8 mg). Offline 2D R-HSCCC with extrusion mode was used to separate Fr.2, and astilbin (4, 37.3 mg), neoisoastilbin (5, 8.8 mg), engeletin (7, 7.9 mg), and a mixture of two polyphenols were obtained from 100 mg of Fr.2. The mixture of two polyphenols was further separated by pre-HPLC, yielding neoastilbin (3, 15.2 mg) and isoastilbin (6, 9.9 mg). The purities of these seven compounds were all over 96.0%. Their structures were identified by MS and NMR. The results demonstrated that the strategy based on offline 2D R-HSCCC with extrusion mode was a powerful tool to separate the main compounds from the rhizome of Smilax glabra and valued for the preparative separation compounds with broad K-values and similar structures.

Semi-Preparative Separation of 10 Caffeoylquinic Acid Derivatives Using High Speed Counter-Current Chromatogaphy Combined with Semi-Preparative HPLC from the Roots of Burdock (Arctium lappa L.).

Burdock roots are healthy dietary supplements and a kind of famous traditional Chinese medicine, which contains large amounts of caffeoylquinic acid derivatives. However, little research has been reported on the preparative separation of these compounds from burdock roots. In the present study, a combinative method of HSCCC and semi-preparative HPLC was developed for the semi-preparative separation of caffeoylquinic acid derivatives from the burdock roots. The ethyl acetate extract of burdock roots was first fractionated by MCI macroporous resin chromatography and give three fractions (Fr. 1-3) from the elution of 40% methanol. Then, these three fractions (120 mg) were separately subjected to HSCCC for purification with the solvent system composed of petroleum ether-ethyl acetate-methanol-water at different volume ratios, and the mixtures were further purified by semi-preparative HPLC. As a result, a total of eight known caffeoylquinic acid derivatives including 3-O-caffeoylquinic acid (32.7 mg, 95.7%), 1,5-O- dicaffeoylquinic acid (4.3 mg, 97.2%), 3-O-caffeoylquinic acid methyl ester (12.1 mg, 93.2%), 1,3-O-dicaffeoylquinic acid (42.9 mg, 91.1%), 1,5-O-dicaffeoyl-3-O-(4-maloyl)-quinic acid (4.3 mg, 84.5%), 4,5-O-dicaffeoylquinic acid (5.3 mg, 95.5%), 1,5-O-dicaffeoyl-3-O-succinylquinic acid (8.7 mg, 93.4%), and 1,5-O-dicaffeoyl-4-O-succinylquinic acid (1.7 mg, 91.8%), and two new compounds were obtained. The new compounds were 1,4-O-dicaffeoyl-3-succinyl methyl ester quinic acid (14.6 mg, 96.1%) and 1,5-O-dicaffeoyl-3-O-succinyl methyl ester quinic acid (3.1 mg, 92.6%), respectively. The research indicated that the combination of HSCCC and semi-preparative HPLC is a highly efficient approach for preparative separation of the instability and bioactive caffeoylquinic acid derivatives from natural products.

Chemical Constituents of the Ethyl Acetate Extract from Diaphragma juglandis Fructus and Their Inhibitory Activity on Nitric Oxide Production In Vitro.

Diaphragma juglandis fructus contains various bioactive constituents. Fourteen compounds were isolated from Diaphragma juglandis fructus by preparative high performance liquid chromatography (pre-HPLC) and high-speed counter-current chromatography (HSCCC). Their structures were identified by nuclear magnetic resonance (NMR) and electrospray ionization mass spectrometry (ESI-MS). Compounds (+)-dehydrovomifoliol (12), (6R,9R)-9-hydroxymegastigman-4-en-3-one (13) and (6R,9S)-9-hydroxymegastigman-4-en-3-one (14) are found from Juglans regia L. for the first time. Compounds dihydrophaseic acid (2), blumenol B (3) and (4S)-4-hydroxy-1-tetralone (11) are isolated from Diaphragma juglandis fructus for the first time. The anti-inflammatory effects of isolated compounds were evaluated by an in vitro model of lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Compounds gallic acid (1), ethyl gallate (9) and (+)-dehydrovomifoliol (12) exhibited inhibitory activity on the nitric oxide production of RAW 264.7 at a concentration of 25 µM. The result indicated that the combination HSCCC with pre-HPLC is an effective way for compound separation and purification. And Diaphragma juglandis fructus constituents have the potential for the treatment of inflammatory-related diseases.

A Strategy for Preparative Separation of 10 Lignans from Justicia procumbens L. by High-Speed Counter-Current Chromatography.

Ten compounds, including three lignan glycosides and seven lignans, were purified from Justicia procumbens L. in 8 h using an efficient strategy based on high-speed counter-current chromatography (HSCCC). The two-phase solvent system composed of petroleum-ethyl acetate-methanol-H2O (1:0.7:1:0.7, v/v) was firstly employed to separate the crude extract (320 mg), from which 19.3 mg of justicidin B (f), 10.8 mg of justicidin A (g), 13.9 mg of 6'-hydroxyjusticidin C (h), 7.7 mg of justicidin E (i), 6.3 mg of lignan J1 (j) were obtained with 91.3 mg of enriched mixture of compounds a-e. The enriched mixture (91.3 mg) was further separated using the solvent system consisting of petroleum-ethyl acetate-methanol-H2O (3:3.8:3:3.8, v/v), yielding 12.1 mg of procumbenoside E (a); 7.6 mg of diphyllin-1-O-ß-d-apiofuranoside (b); 7.4 mg of diphyllin (c); 8.3 mg of 6'-hydroxy justicidin B (d); and 7.9 mg of diphyllin acetyl apioside (e). The purities of the 10 components were all above 94%, and their structures were identified by NMR and ESI-MS spectra. The results demonstrated that the strategy based on HSCCC for the separation of lignans and their glycosides was efficient and rapid.

[Optimization of Aqueous Two-Phase Extraction of Polysaccharide from Schisandra chinensis Using Response Surface Methodology with Box-Behnken Design].

Objective: To optimize the extraction condition of polysaccharide from Schisandra chinensis. Aqueous two-phase extraction（ ATPE） method was used, based on Box-Behnken design with Response surface methodology（ BBD-RSM）. Methods: solvent volume,K2HPO4 and PEG6000 were selected as the investigation factors by the single-factor experiment, and the overall desirability（ OD）of phase volume ratio, partition coefficient and extraction rates were the reponse value. BBD-RSM was used to optimize the extraction process. . Results: The optimal parameters were as follows,the solvent volume was 5 m L,the addition amount of K2HPO4 was 1. 0 g, the addition amount of PEG6000 was 1. 8 g and centrifugation time was 9 min, which indicated that the model had a good predictability. The predicted value was 0. 950,and the deviation between observed and predicted values was 3. 94% Conclusion: The ATPE technology is easy to operate and cost-effective for the extraction of polysaccharides from Schisandra chinensis.

Enrichment and separation of antitumor triterpene acids from the epidermis of Poria cocos by pH-zone-refining counter-current chromatography and conventional high-speed counter-current chromatography.

Triterpene acids were extracted from the epidermis of Poria cocos (Schw.) Wolf. These acids were found to inhibit the growth of lung cancer cells in vitro and in vivo. An efficient method for the preparative separation of antitumor triterpene acids was established that involves the combination of pH-zone-refining counter-current chromatography and conventional high-speed counter-current chromatography. We used pH-zone-refining counter-current chromatography to concentrate the triterpene acids using a two-phase solvent system composed of petroleum ether/ethyl acetate/methanol/water (3:7:5:5, v/v/v/v), trifluoroacetic acid (10 mM) was added to the upper phase as a retainer, and ammonia (10 mM) was added to the lower phase as an eluter. As a result, 200 mg concentrate of triterpene acids was obtained from 1.0 g of crude extract. The concentrate was further separated by conventional high-speed counter-current chromatography using a solvent system composed of petroleum ether/ethyl acetate/methanol/water (0.8:1.2:1.2:0.9, v/v), yielding 50 mg of poricoic acid A and 5 mg of poricoic acid B from 120 mg concentrate, respectively. The inhibitory activity of the major compound on lung A549 cells was examined and poricoic acid A was found to significantly inhibit the growth of A 549 cells.

Identification of key anti-neuroinflammatory components in Gastrodiae Rhizoma based on spectrum-effect relationships and its mechanism exploration.

Gastrodiae Rhizoma was proven to have anti-inflammatory activity based on its main component of 4-hydroxybenzyl alcohol (4-HBA) and gastrodin (GAS). However, the anti-inflammatory activity of other phenols has been less reported. In this study, the n-BuOH extract was selected as the active anti-inflammatory part of Gastrodiae Rhizoma based on the LPS-induced inflammatory BV-2 cells. The spectral-effect relationship analysis of the n-BuOH extract showed the main effective components were GAS, 4-HBA, parishin A (PA), parishin B (PB), and parishin C (PC). Among them, PB could reduce LPS-induced expression of nitric oxide (NO), intracellular ROS, TNF-α, IL-6, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). Molecular docking predicted that PB had a good binding capacity to AMPKα and SIRT1 proteins of -12.1 kJ/mol and -7.6 kJ/mol, respectively. The Western Blot results further demonstrated that PB could inhibit NF-κB pathway by activating AMPK/SIRT1 pathway, thus exerting anti-LPS-induced neuroinflammatory effects. This study provides a referable idea for solving the problem of unclear action of TCM with complex compositions and is of great significance for the development of innovative medicines of traditional Chinese medicine.

Screening and characterization of potential anti-gout components from Polygonum cuspidatum by integration off-line two-dimensional liquid chromatography-mass spectrometry with affinity ultrafiltration and on-line HPLC-ABTS.

Polygonum cuspidatum (P. cuspidatum) is a traditional herbal medicine with a long history and proven efficacy in treating gout. However, due to the complexity of composition and extensive content distribution, the substance basis of its anti-gout effectiveness is still unclear. A strategy was proposed via integrating off-line two-dimensional liquid chromatography (2D-LC) and targeted rapid screening technology based on ultrafiltration-liquid chromatography-mass spectrometry (UF-LC/MS) and on-line high-performance liquid chromatography-2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (HPLC-ABTS) to accomplish high coverage and high throughput screening of anti-gout components from P. cuspidatum. As a result, twenty components were screened from P. cuspidatum extract with both xanthine oxidase (XOD) inhibitory activity and free radical scavenging activity, then were preliminarily identified by high-resolution electrospray ionization-quadrupole-time-of-flight mass spectrometer (ESI-Q-TOF/MS). The screened results were verified by the in vitro assays. Meanwhile, molecular docking further elucidated that the screened bioactive ingredients had favourable binding capabilities with XOD. The performance of this study can achieve high efficiency and high coverage screening of the anti-gout components from P. cuspidatum, which provides methodology and strategy support for the rapid screening of bioactive ingredients from complex medicinal plants.

Prediction of quality markers in Maren Runchang pill for constipation using machine learning and network pharmacology.

Maren Runchang pill (MRRCP) is a Chinese patent medicine used to treat constipation in clinics. It has multi-component and multi-target characteristics, and there is an urgent need to screen markers to ensure its quality. The aim of this study was to screen quality markers of MRRCP based on a "differential compounds-bioactivity" strategy using machine learning and network pharmacology to ensure the effectiveness and stability of MRRCP. In this study, UPLC-Q-TOF-MS/MS was used to identify chemical compounds in MRRCP and machine learning algorithms were applied to screen differential compounds. The quality markers were further screened by network pharmacology. Meanwhile, molecular docking was used to verify the screening results of machine learning and network pharmacology. A total of 28 constituents in MRRCP were identified, and four differential compounds were screened by machine learning algorithms. Subsequently, a total of two quality markers (rutin and rubiadin) in MRRCP. Additionally, the molecular docking results showed that quality markers could spontaneously bind to core targets. This study provides a reference for improving the quality evaluation method of MRRCP to ensure its quality. More importantly, it provided a new approach to screen quality markers in Chinese patent medicines.

Structure and properties of acidic polysaccharides isolated from Massa Medicata Fermentata: Neuroprotective and antioxidant activity.

Massa Medicata Fermentata (MMF) is a fermented food with therapeutic effects. Previous studies suggested that after stir-frying, the uronic acid content in MMF crude polysaccharides increases, and the pH value decreases, which is caused by the change in acidic polysaccharides. However, the detailed physicochemical properties and structure-activity correlation of the acidic polysaccharides in MMF have not been fully explored. In this study, two acidic polysaccharides (SMMFAP and CMMFAP) were isolated from the MMF and its stir-fried product, respectively. Their structural characteristics and bioactivities were comparatively studied, and the structure-activity correlation was examined. Our findings revealed that the SMMFAP had a higher average Mw and higher Gal and Man content than the CMMFAP. Both the SMMFAP and CMMFAP were mainly composed of Xyl, Man, and Gal residues, whereas the CMMFAP had fewer linkage types. Additionally, the CMMFAP exhibited stronger neuroprotective activity than the SMMFAP owing to its higher content of 1,6-linked-Galp, while the SMMFAP exhibited better antioxidant activity, which might be related to its higher average Mw. Our findings suggest that acidic polysaccharides may be the active substances that cause differences in effectiveness between the sheng and chao MMF. Furthermore, the research qualified the SMMFAP and CMMFAP with different potential applications.