A Rapid and Accurate UHPLC Method for Determination of Monosaccharides in Polysaccharides of Different Sources of Radix Astragali and Its Immune Activity Analysis.

With the escalating demand for Astragalus polysaccharides products developed from Radix Astragali (RA), the necessity for quality control of polysaccharides in RA has become increasingly urgent. In this study, a specific method for the simultaneous determination of seven monosaccharides in polysaccharides extracted from Radix Astragali (RA) has been developed and validated using ultra-performance liquid chromatography equipped with an ultraviolet detector (UHPLC-UV) for the first time. The 1-phenyl-3-methyl-5-pyrazolone (PMP) derivatizations were separated on a C18 column (Waters ACQUITYTM, Milfor, MA, USA, 1.8 µm, 2.1 × 100 mm) using gradient elution with a binary system of 5 mm ammonium formate (0.1% formic acid)-acetonitrile for 24 min. Additionally, seven monosaccharides showed good linear relationships (R2, 0.9971-0.9995), adequate precision (RSD < 4.21%), and high recoveries (RSD < 4.70%). The established method was used to analyze 109 batches of RA. Results showed that the Astragalus polysaccharides (APSs) mainly consist of mannose (Man), rhamnose (Rha), glucose (Glu), galactose (Gal), arabinose (Ara), xylose (Xyl); and fucose (Fuc); however, their composition was different among RA samples from different growth patterns, species, growth years, and origins, and the growth mode of RA and the age of wild-simulated RA can be accurately distinguished by principal component analysis (PCA). In addition, the immunological activity of APSs were also evaluated jointly by measurement of the NO release with RAW264.7, with the results showing that APSs have a promoting effect on the release of NO and exhibit a significant correlation with Man, Glu, Xyl, and Fuc contents. Accordingly, the new established monosaccharides analytical method and APS-immune activity determination in this study can provide a reference for quality evaluation and the establishment of quality standards for RA.

Classification and Authentication of Lonicerae Japonicae Flos and Lonicerae Flos by Using 1H-NMR Spectroscopy and Chemical Pattern Recognition Analysis.

Lonicerae japonicae flos and Lonicerae flos are increasingly widely used in food and traditional medicine products around the world. Due to their high demand and similar appearance, they are often used in a confused or adulterated way; therefore, a rapid and comprehensive analytical method is highly required. In this case, the comparative analysis of a total of 100 samples with different species, growth modes, and processing methods was carried out by nuclear magnetic resonance (1H-NMR) spectroscopy and chemical pattern recognition analysis. The obtained 1H-NMR spectrums were employed by principal component analysis (PCA), partial least-squares discriminant analysis (PLS-DA), orthogonal partial least-squares discriminant analysis (OPLS-DA), and linear discriminant analysis (LDA). Specifically, after the dimensionality reduction of data, linear discriminant analysis (LDA) exhibited good classification abilities for the species, growth modes, and processing methods. It is worth noting that the sample prediction accuracy from the testing set and the cross-validation predictions of the LDA models were higher than 95.65% and 98.1%, respectively. In addition, the results showed that macranthoidin A, macranthoidin B, and dipsacoside B could be considered as the main differential components of Lonicerae japonicae flos and Lonicerae Flos, while secoxyloganin, secologanoside, and sweroside could be responsible for distinguishing cultivated and wild Lonicerae japonicae Flos. Accordingly, 1H-NMR spectroscopy combined with chemical pattern recognition gives a comprehensive overview and provides new insight into the quality control and evaluation of Lonicerae japonicae flos.

Discrimination of Radix Astragali from Different Growth Patterns, Origins, Species, and Growth Years by an H1-NMR Spectrogram of Polysaccharide Analysis Combined with Chemical Pattern Recognition and Determination of Its Polysaccharide Content and Immunological Activity.

The fraud phenomenon is currently widespread in the traditional Chinese medicine Radix Astragali (RA) market, especially where high-quality RA is substituted with low-quality RA. In this case, focused on polysaccharides from RA, the classification models were established for discrimination of RA from different growth patterns, origins, species, and growth years. 1H Nuclear Magnetic Resonance (H1-NMR) was used to establish the spectroscopy of polysaccharides from RA, which were used to distinguish RA via chemical pattern recognition methods. Specifically, orthogonal partial least squares discriminant analysis (OPLS-DA) and linear discriminant analysis (LDA) were used to successfully establish the classification models for RA from different growth patterns, origins, species, and growth years. The satisfactory parameters and high accuracy of internal and external verification of each model exhibited the reliable and good prediction ability of the developed models. In addition, the polysaccharide content and immunological activity were also tested, which was evaluated by the phagocytic activity of RAW 264.7. And the result showed that growth patterns and origins significantly affected the quality of RA. However, there was no significant difference in the aspects of origins and growth years. Accordingly, the developed strategy combined with chemical information, biological activity, and multivariate statistical method can provide new insight for the quality evaluation of traditional Chinese medicine.

Arg-liposome-amplified colorimetric immunoassay for selective and sensitive detection of cystatin C to predict acute kidney injury.

Cystatin C (Cys C) has been considered as a novel biomarker of kidney disease, which is thought to be a better indicator of glomerular filtration rate than creatinine (Scr) in the prediction of acute kidney injury (AKI). Hence, there is strong need to develop a precise, rapid and simple detection method for Cys C. Here we reported a Arg-liposome-amplified colorimetric immunoassay for the detection of Cys C to predict AKI. Cys C antibodies are conjugated on the surface of magnetic beads (MBs) and arginine (Arg)-loaded liposomes to form Ab1-MBs and Ab2-Arg-liposomes, respectively. When Ab1-MBs captured Cys C, Ab2-Arg-liposomes are added and incubated to form the immuno-sandwich complex. After magnetic separation, the surfactant Triton ×100 is added to damage the liposomes, leading to the release of Arg which can induce the gold nanoparticles aggregation. Therefore, the discoloration can be used for visual and quantitative detection of Cys C. Notably, the method has a linear relation in the range of 10-100 µg/L for Cys C with a limit of detection 4.32 µg/L, which is lower than some of the previous reports. In addition, the AKI mice serum samples were tested by the developed method, which were in good agreement with ELISA results. More intriguingly, the results of cisplatin induced acute kidney injury in mice showed that the method could be used to evaluate the protective effect of astragalus membranaceus (AM) on AKI by detecting Cys C in serum, providing a new strategy for screening renal protective drugs. Accordingly, a rapid and highly sensitive Cys C detection system was established with great potential for clinical diagnostics.

Comparison of the Chemical Components, Efficacy and Mechanisms of Action of Chrysanthemum morifolium Flower and Its Wild Relative Chrysanthemum indicum Flower against Liver-Fire Hyperactivity Syndrome of Hypertension via Integrative Analyses.

To clarify the differences in the clinical application scope of Chrysanthemum morifolium flower (CMF) and Chrysanthemum indicum flower (CIF), two herbs of similar origin, an integrated strategy of network pharmacology, molecular pharmacology, and metabolomics was employed, with a view to investigating the commonalities and dissimilarities in chemical components, efficacy and mechanisms of action. Initial HPLC-Q-TOF-MS analysis revealed that CMF and CIF had different flavonoid constituents. The biological processes underlying the therapeutic effects of CMF and CIF on liver-fire hyperactivity syndrome of hypertension (LFHSH) were predicted to be related to inflammatory response, fatty acid production, and other pathways based on network pharmacology analysis. ELISA, molecular docking, Western blot, and metabolomics techniques showed similar effects of CMF and CIF in lowering blood pressure, resistance to tissue, organ and functional damage, and dyslipidemia. However, distinct effects were found in the regulation of inflammatory response, PI3K-Akt and NF-κB signaling pathways, lipid anabolism, renin-angiotensin system, and metabolic abnormalities. The comparable efficacies of CMF and CIF, despite having distinct mechanisms of action, may be attributed to the integration and counteraction of their different regulating capabilities on the above anti-LFHSH mechanisms. This study offers a vital platform for assessment of differential and precise applications of herbs of close origin with similar but slightly different medicinal properties, and provides a research strategy for bridging Chinese medicine and modern precision medicine.

A nano-preparation approach to enable the delivery of daphnoretin to potentiate the therapeutical efficacy in hepatocellular cancer.

Daphnoretin (DAP), isolated from a traditional Chinese medicine Wikstroemia indica (Linn. C. A. Meyer), could induce apoptosis of hepatocellular cancer (HCC) and inhibit tumor growth. However, the application of DAP in cancer therapies was hampered because to its poor solubility. Herein, this study aimed to design an approach of double-targeted nano-preparation to enable the delivery of DAP to potentiate the therapeutical efficacy in liver cancer via glycyrrhetinic acid-polyethylene glycol-block-poly (D,L-lactic acid)/polyethylene glycol-block-poly (D,L-lactic acid)-DAP (GPP/PP-DAP). In particular, the purity of separated DAP was up to 98.12% for preparation research. GPP/PP-DAP was successfully prepared by the thin-film hydration method. Subsequently, the GPP/PP-DAP was optimized by univariate analysis and the response surface methodology, producing a stable and systemically injectable nano-preparation. Impressively, on the one hand, cytotoxicity studies showed that the IC50 of the GPP/PP-DAP was lower than that of free DAP. On the other hand, the GPP/PP-DAP was more likely to be endocytosed by HepG2 cells and targeted to the liver with orthotopic tumors, potentiating the therapeutical efficacy in HCC. Collectively, both in vitro and in vivo results indicated the excellent tumor inhibition and liver targeting of GPP/PP-DAP, suggesting the nano-preparation could serve as a potential drug delivery system for natural ingredients with anti-hepatoma activity to lay the theoretical foundation for clinical application.

Rapid Fluorescence Sensor Guided Detection of Urinary Tract Bacterial Infections.

Introduction: Urinary tract infections (UTI) are one of the most serious human bacterial infections affecting millions of people every year. Therefore, simple and reliable identification of the urinary tract pathogenic bacteria within a few minutes would be of great significance for diagnosis and treatment of clinical patients with UTIs. In this study, the fluorescence sensor was reported to guide the detection of urinary tract bacterial infections rapidly. Methods: The Ami-AuNPs-DNAs sensor was fabricated by the amino-modified Au nanoparticles (Ami-AuNPs) and six DNAs signal molecules, which bound to the urinary tract pathogenic bacteria and generated corresponding response signals. Further, based on the collected response signals, identification was performed by principal component analysis (PCA) and linear discriminant analysis (LDA). The Ami-AuNPs and Ami-AuNPs-DNAs were characterized by transmission electron microscopy, UV-vis absorption spectrum, Fourier transform infrared spectrum, dynamic light scattering and zeta potentials. Thereafter, the Ami-AuNPs-DNAs sensor was used to discriminate and identify five kinds of urinary tract pathogenic bacteria. Moreover, the quantitative analysis performance towards individual bacteria at different concentrations were also evaluated. Results: The Ami-AuNPs-DNAs sensor were synthesized successfully in terms of spherical, well-dispersed and uniform in size, which could well discriminate five main urinary tract pathogenic bacteria with unique fingerprint-like patterns and was sufficiently sensitive to determine individual bacteria with a detection limit to 1×107 cfu/mL. Furthermore, the sensor had also been successfully applied to identify bacteria in urine samples collected from clinical UTIs. Conclusion: The developed fluorescence sensor could be applied to rapid and accurate discrimination of urinary tract pathogenic bacteria and holds great promise for the diagnosis of the disease caused by bacterial infection.

Integrating Anti-Influenza Virus Activity and Chemical Pattern Recognition to Explore the Quality Evaluation Method of Lonicerae Japonicae Flos.

Lonicerae japonicae flos (LJF, Lonicera japonica Thunb.) is adopted as a core herb for preventing and treating influenza. However, the anti-influenza virus components of LJF and the impact of quality-affecting factors on the anti-influenza activity of LJF have not been systematically investigated. In this study, a strategy integrating anti-influenza virus activity, ultrahigh-performance liquid chromatography fingerprint and chemical pattern recognition was proposed for the efficacy and quality evaluation of LJF. As a result, six bioactive compounds were screened out and identified as neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, 4,5-Di-O-caffeoylquinic acid, sweroside and secoxyloganin. Based on the bioactive compounds, chemical pattern recognition models of LJF were established by a linear discriminant analysis (LDA). The results of the LDA models and anti-influenza virus activity demonstrated that cultivation pattern significantly affected the anti-influenza effect of LJF and that the neuraminidase inhibition rate of wild LJF was significantly higher than that of cultivated LJF. Moreover, the quality of LJF samples with different processing methods and geographical origins showed no obvious difference. Overall, the proposed strategy in the current study revealed the anti-influenza virus components of LJF and provided a feasible method for thequality evaluation of LJF, which has great importance for assuring the clinical effect against influenza of LJF.

Discrimination of Lonicerae Japonicae Flos according to species, growth mode, processing method, and geographical origin with ultra-high performance liquid chromatography analysis and chemical pattern recognition.

Lonicerae japonicae flos (LJF, Lonicera japonica Thunb.) is often confused and/or adulterated with Lonicerae flos (LF, Lonicera macrantha (D.Don) Spreng.). Ecological conditions and processing methods strongly influenced the safety and efficacy of LJF. For the strict quality control of LJF, a rapid and feasible strategy for identification and classification of LJF by species, growth mode, processing method and geographical origin, based on chromatographic profiles and pattern recognition analysis, in 119 batches of Lonicera samples was systematically established. Firstly, comprehensive analysis of the chemical compositions of LJF was achieved using ultra-high performance liquid chromatography (UHPLC). Next, unsupervised principal component analysis showed that the influence of species, growth mode, processing method and geographical origin displayed a decreasing trend. Subsequently, classification models for authentication of LJF samples were established by linear discriminant analysis (LDA) with good classification abilities. Finally, sweroside and secoxyloganin could be considered as markers associated of cultivated and wild LJF, respectively, while 3-O-caffeoylquinic acid and 3,5-Di-O-caffeoylquinic acid could be regarded as markers for LF. Consequently, the findings suggest that UHPLC profiles combined with pattern recognition analysis is precise and feasible strategy for the discrimination and quality control of LJF.

Chemical characteristics of Rhodiola Crenulata and its mechanism in acute mountain sickness using UHPLC-Q-TOF-MS/MS combined with network pharmacology analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Rhodiola crenulata (Hook.f. & Thomson) H.Ohba has a long history of clinical application for the prevention and treatment of acute mountain sickness (AMS) in traditional Chinese medicine. However, gaps in knowledge still exist in understanding the underlying mechanisms of Rhodiola crenulata against AMS. AIMS: To address this problem, a comprehensive method was established by combining UHPLC-Q-TOF-MS/MS analysis and network pharmacology. MATERIALS AND METHODS: The ingredients of Rhodiola crenulata were comprehensively analyzed using UHPLC-Q-TOF-MS/MS method. On this basis, a network pharmacology method incorporated target prediction, protein-protein interaction network, gene enrichment analysis and components-targets-pathways network was performed. Finally, the possible mechanisms were verified through molecular docking, in vitro and in vivo experiments. RESULTS: A total of 106 constituents of Rhodiola crenulata were charactered via UHPLC-Q-TOF-MS/MS. The 98 potentially active compounds out of 106 were screened and corresponded to 53 anti-AMS targets. Gene enrichment analysis revealed that hypoxia and inflammation related genes may be the central factors for Rhodiola crenulata to modulate AMS. Molecular docking revealed that TNF, VEGFA and HIF-1α had high affinities to Rhodiola crenulata compounds. Subsequently, Rhodiola crenulata extract was indicated to inhibit the protein expression level of TNF in hypoxia induced H9c2 cells. Lastly, Rhodiola crenulata extract was further verified to ameliorate heart injury and decreased the heart levels of TNF, VEGFA and HIF-1α in acute hypoxia-induced rats. CONCLUSIONS: This study used UHPLC-Q-TOF-MS/MS analysis and a network pharmacology to provide an important reference for revealing the potential mechanism of Rhodiola crenulata in the prevention and treatment of AMS.

The therapeutic effect of Xuanbai Chengqi Decoction on chronic obstructive pulmonary disease with excessive heat in the lung and fu-organs based on gut and lung microbiota as well as metabolic profiles.

As a prescription for treating lung inflammation and intestinal diseases, Xuanbai Chengqi Decoction (XBCQD) in clinical practice can effectively treat COPD with excessive heat in the lung and fu-organs, which is characterized by phlegm-heat accumulation in the lung and constipation. This study aims to find the potential biomarkers of COPD with excessive heat in the lung and fu-organs from two aspects of lung and intestine based on metabolomics and microbiota analysis, and to evaluate the efficacy of XBCQD as well as to explore the mechanism of drug function according the regulating effect of drugs on these markers. The HPLC-Q-TOF-MS/MS, 16SrDNA technology and multiple statistical methods were used to trace the process of disease and curative effect with XBCQD. Results showed that the onset and development of disease was associated with the imbalance of 41 differential metabolites in plasma, bronchoalveolar lavage fluid and feces and 82 bacteria at the levels of phylum, class, order, family and genus from lung and intestine, including Escherichia-Shigella. However, after treatment with XBCQD, 30 differential metabolites mainly involving in the metabolism of linoleic acid, taurine and hypotaurine metabolism, arachidonic acid metabolism, biosynthesis of primary bile acids, tryptophan metabolism, arginine and proline metabolism and 65 pulmonary and intestinal bacteria at all levels were reversed in the drug group. In addition, the results of the correlation analysis showed that specific microbiota from lung and intestine and reversed differential metabolites had a significant correlation, and they could affect each other in the course of disease occurrence and treatment. This study preliminarily confirmed that XBCQD can be used to treat COPD with excessive heat in the lung and fu-organs through lung-intestine simultaneous treatment. It also provided new strategies for the treatment of lung diseases or intestinal diseases, and new research ideas for the evaluation of drug efficacy.

Leonurus japonicus Houtt. (Motherwort): Systematic research through chemical profiling, stability under controlled conditions and pharmacokinetic analysis on screening Q-markers for quality control.

Leonurus japonicus Houtt. (Motherwort) is the fresh or dried aerial part of Leonurus japonicus Houtt. (Labiaceae), which is widely used in clinical practice and daily life, used to treat gynecological diseases. However, the differences between different parts, single index component in Pharmacopoeias and the less stability of active ingredients affect its clinical efficacy. This study aimed to find the multi-active compounds between different parts of Motherwort to ensure its clinical efficacy, which related to stability and had pharmacokinetic behavior. Firstly, HPLC-Q-TOF-MS/MS was used to analyze the components in vitro and in vivo, as well as multivariate statistical analysis and network pharmacology analysis was conducted to find the significant different components related to activity. Secondly, the content determination methods were established to study the stability of effective components during storage in order to establish the content limit for quality control of Motherwort. Thirdly, UFLC-MS/MS was used to analyze the pharmacokinetic behavior of active components in Motherwort. The results showed that a total of 131 chemical constituents were identified in vitro and 21 prototype absorption compounds and 72 metabolites were found in vivo. Meantime, multivariate statistical analysis and network pharmacology analysis was combined to find that leonurine, stachydrine and trigonelline were activity-related substance, which could be used as active components related to pharmacodynamics in different parts. Then the stability variation trend and content limit of three alkaloids were found, which could be used for the quality control of Motherwort. Furthermore, the results showed that three alkaloids had pharmacokinetic behavior in vivo. 3 alkaloids were screened, which could be used as active components most closely related to pharmacodynamics among different parts. The stable stage, assay tolerance and pharmacokinetic characteristics were studied by the active substances, which could provide a basis for quality control and clinical medication of Motherwort.

The Study of Steaming Durations and Temperatures on the Chemical Characterization, Neuroprotective, and Antioxidant Activities of Panax notoginseng.

Panax notoginseng (PN) is one of the most valuable traditional Chinese medicines and has extensive pharmacological effects. Recent studies demonstrated that PN exhibited pharmacological effect related to Alzheimer's disease (AD). However, whether steaming process can boost its anti-AD activity is still unexplored. To fill this gap, effects of steaming durations and temperatures on the chemical characterization, neuroprotective and antioxidant activities of PN were systematically investigated in this study. HPLC fingerprint coupled with quantitative analysis demonstrated striking conversion of original saponins to less polar ones with the increase in the steaming time and temperature. In the viewpoint of anti-AD activity on neuroprotective and antioxidant effects, several steamed PN samples (110°C-6/8/10 h, 120°C -4/6 h samples) displayed a significant increase both in cell viability and oxygen radical absorption capacity (ORAC) values compared with the no steamed one (P < 0.01 or P < 0.005). Steaming temperature had the greater impact on the change of chemical composition and anti-AD activity of PN. Moreover, the spectrum-effect relationship analysis revealed that the transformed saponins were partially responsible for the increased neuroprotective and antioxidant effects of steamed PN. Therefore, steamed PN could be used as a potential crude drug for prevention and treatment of AD.

Application of UHPLC Fingerprints Combined with Chemical Pattern Recognition Analysis in the Differentiation of Six Rhodiola Species.

Rhodiola, especially Rhodiola crenulate and Rhodiola rosea, is an increasingly widely used traditional medicine or dietary supplement in Asian and western countries. Because of the phytochemical diversity and difference of therapeutic efficacy among Rhodiola species, it is crucial to accurately identify them. In this study, a simple and efficient method of the classification of Rhodiola crenulate, Rhodiola rosea, and their confusable species (Rhodiola serrata, Rhodiola yunnanensis, Rhodiola kirilowii and Rhodiola fastigiate) was established by UHPLC fingerprints combined with chemical pattern recognition analysis. The results showed that similarity analysis and principal component analysis (PCA) could not achieve accurate classification among the six Rhodiola species. Linear discriminant analysis (LDA) combined with stepwise feature selection exhibited effective discrimination. Seven characteristic peaks that are responsible for accurate classification were selected, and their distinguishing ability was successfully verified by partial least-squares discriminant analysis (PLS-DA) and orthogonal partial least-squares discriminant analysis (OPLS-DA), respectively. Finally, the components of these seven characteristic peaks were identified as 1-(2-Hydroxy-2-methylbutanoate) ß-D-glucopyranose, 4-O-glucosyl-p-coumaric acid, salidroside, epigallocatechin, 1,2,3,4,6-pentagalloyglucose, epigallocatechin gallate, and (+)-isolarisiresinol-4'-O-ß-D-glucopyranoside or (+)-isolarisiresinol-4-O-ß-D-glucopyranoside, respectively. The results obtained in our study provided useful information for authenticity identification and classification of Rhodiola species.

Based on Multi-Activity Integrated Strategy to Screening, Characterization and Quantification of Bioactive Compounds from Red Wine.

According to French Paradox, red wine was famous for the potential effects on coronary heart disease (CHD), but the specific compounds against CHD were unclear. Therefore, screening and characterization of bioactive compounds from red wine was extremely necessary. In this paper, the multi-activity integrated strategy was developed and validated to screen, identify and quantify active compounds from red wine by using ultra high performance liquid chromatography-fraction collector (UHPLC-FC), ultra fast liquid chromatography-quadrupole-time-of-flight/mass spectrometry (UFLC-Q-TOF/MS) and bioactive analysis. UHPLC-FC was employed to separate and collect the components from red wine, which was further identified by UFLC-Q-TOF/MS to acquire their structural information. Furthermore, the active fractions were tested for antioxidant activity, inhibitory activity against thrombin and lipase activities in vitro by the activity screening kit. As the results, there were 37 fractions had antioxidant activity, 22 fractions had thrombin inhibitory activity and 28 fractions had lipase inhibitory activity. Finally, 77 active components from red wine were screened and 12 ingredients out of them were selected for quantification based on the integration of multi-activity. Collectively, the multi-activity integrated strategy was helpful for the rapid and effective discovery of bioactive components, which provided reference for exploring the health care function of food.

Physiologically Based Pharmacokinetic Models Are Effective Support for Pediatric Drug Development.

Pediatric drug development faces many difficulties. Traditionally, pediatric drug doses are simply calculated linearly based on the body weight, age, and body surface area of adults. Due to the ontogeny of children, this simple linear scaling may lead to drug overdose in pediatric patients. The physiologically based pharmacokinetic (PBPK) model, as a mathematical model, contributes to the research and development of pediatric drugs. An example of a PBPK model guiding drug dose selection in pediatrics has emerged and has been approved by the relevant regulatory agencies. In this review, we discuss the principle of the PBPK model, emphasize the necessity of establishing a pediatric PBPK model, introduce the absorption, distribution, metabolism, and excretion of the pediatric PBPK model, and understand the various applications and related prospects of the pediatric PBPK model.

Developing a novel and simple biosensor for Cystatin C as a fascinating marker of glomerular filtration rate with DNase I-aided recycling amplification strategy.

Cystatin C (Cys C) has been proposed as a fascinating glomerular filtration rate (GFR) marker for early detection of acute kidney injury and chronic kidney disease. However, most of traditional methods for Cys C detection are immunoassays, which was tedious to perform and unfriendly for economics. In this work, a novel and simple biosensor for the sensitive measurement of Cys C via DNase I-aided recycling amplification strategy was successfully constructed based on the graphene oxide (GO) and fluorophore-labelled aptamer, which can be used to the early prediction of kidney injury. The fluorescence of fluorophore-labelled aptamer was quenched by GO based on the Fluorescence Resonance Energy Transfer (FRET) and recovered with the existence of Cys C. In addition, the DNase I enzyme would digest the fluorophore-labelled aptamer and dissociate the Cys C to launch the next reaction, resulting in an increase of signal amplification. Hence, the limit of detection is found to be 0.16 ng mL-1, which is almost 3 times lower than that without DNase I. Consequently, the developed biosensor offers a novel approach towards simple and rapid detection of Cys C based on the integration of GO and aptamer. Conceivably, this strategy holds a wide scope in the application of numerous other analytes if corresponding aptamers are available.

Fast analysis of alkaloids from different parts of Mahonia bealei (Fort.) Carr. studied for their anti-Alzheimer's activity using supercritical fluid chromatography.

In this study, a rapid and highly efficient method was developed for the separation of eight isoquinoline alkaloids using supercritical fluid chromatography. The separation conditions were carefully optimized including stationary phases, additives, backpressure, and temperature. Compared to high-performance liquid chromatography, the use of supercritical fluid chromatography could provide a 13 times faster separation. Subsequently, the method was validated and applied for the determination of eight alkaloids from different parts of Mahonia bealei (Fort.) Carr. (stem, root, leaf, and seed). The results indicated a good repeatability with relative standard deviations for overall precisions lower than 3.2%. The limit of detection was between 0.4 and 2.3 µg/mL while limit of quantitation ranged from 1.5 to 7.5 µg/mL. Recovery ranged from 95.7 to 102.5% indicating a validity of recovery. The content of total eight alkaloids was the highest in stem (66.0 µg/g) and root (65.1 µg/g) compared to leaf or seed. Moreover, anti-acetylcholinesterase activity for those extracts was evaluated by Ellman's colorimetric assay. As a result, the acetylcholinesterase inhibitory activity of the extracted samples was in the following decreasing order: stem > root > leaf or seed. In conclusion, the results indicated that supercritical fluid chromatography could be a useful tool for quality control of Mahonia bealei (Fort.) Carr. containing alkaloids as active compounds.

Chemical Pattern Recognition for Quality Analysis of Lonicerae Japonicae Flos and Lonicerae Flos Based on Ultra-High Performance Liquid Chromatography and Anti-SARS-CoV2 Main Protease Activity.

Lonicerae japonicae flos (L. japonicae flos, Lonicera japonica Thunb.) is one of the most commonly prescribed botanical drugs in the treatment or prevention of corona virus disease 2019. However, L. japonicae flos is often confused or adulterated with Lonicerae flos (L. flos, Lonicera macrantha (D.Don) Spreng., Shanyinhua in Chinese). The anti-SARS-CoV2 activity and related differentiation method of L. japonicae flos and L. flos have not been documented. In this study, we established a chemical pattern recognition model for quality analysis of L. japonicae flos and L. flos based on ultra-high performance liquid chromatography (UHPLC) and anti-SARS-CoV2 activity. Firstly, chemical data of 59 batches of L. japonicae flos and L. flos were obtained by UHPLC, and partial least squares-discriminant analysis was applied to extract the components that lead to classification. Next, anti-SARS-CoV2 activity was measured and bioactive components were acquired by spectrum-effect relationship analysis. Finally, characteristic components were explored by overlapping feature extracted components and bioactive components. Accordingly, eleven characteristic components were successfully selected, identified, quantified and could be recommended as quality control marker. In addition, chemical pattern recognition model based on these eleven components was established to effectively discriminate L. japonicae flos and L. flos. In sum, the demonstrated strategy provided effective and highly feasible tool for quality assessment of natural products, and offer reference for the quality standard setting.

Determination and Comparison of Short-Chain Fatty Acids in Serum and Colon Content Samples: Alzheimer's Disease Rat as a Case Study.

Short-chain fatty acids (SCFAs) are the main microbial fermentation products from dietary fibers in the colon, and it has been speculated that they play a key role in keeping healthy in the whole-body. However, differences in SCFAs concentration in the serum and colon samples had attracted little attention. In this study, we have optimized the extract and analysis methods for the determination of ten SCFAs in both serum and colon content samples. Methanol and acetonitrile were chosen for extraction of SCFAs from serum and colon content samples, respectively. Biological samples were collected from Alzheimer's disease rats treated by extract of Schisandra chinensis (Turcz.) Baill (SC-extract) were taken as research objects. The results showed that, the relative peak intensities of SCFAs in the colon content from all groups were quite similar, and the trend was identical in the serum samples. Compared with the values in humans, the ratio of ten SCFAs in rat's colon was similar, while the percent of acetate in rat's serum was significantly higher. For therapy of Alzheimer's disease (AD), SC-extract decreased the concentration of butyrate, 3-Methyvalerate, and caproate in the serum samples towards the trend of normal rats. This study may help our understanding of how SCFAs are transported across colonic epithelium in healthy and diseased organisms.