A Synergetic Strategy for Brand Characterization of Colla Corii Asini (Ejiao) by LIBS and NIR Combined with Partial Least Squares Discriminant Analysis.

A synergetic strategy was proposed to address the critical issue in the brand characterization of Colla corii asini (Ejiao, CCA), a precious traditional Chinese medicine (TCM). In all brands of CCA, Dong'e Ejiao (DEEJ) is an intangible cultural heritage resource. Seventy-eight CCA samples (including forty DEEJ samples and thirty-eight samples from other different manufacturers) were detected by laser-induced breakdown spectroscopy (LIBS) and near-infrared spectroscopy (NIR). Partial least squares discriminant analysis (PLS-DA) models were built first considering individual techniques separately, and then fusing LIBS and NIR data at low-level. The statistical parameters including classification accuracy, sensitivity, and specificity were calculated to evaluate the PLS-DA model performance. The results demonstrated that two individual techniques show good classification performance, especially the NIR. The PLS-DA model with single NIR spectra pretreated by the multiplicative scatter correction (MSC) method was preferred as excellent discrimination. Though individual spectroscopic data obtained good classification performance. A data fusion strategy was also attempted to merge atomic and molecular information of CCA. Compared to a single data block, data fusion models with SNV and MSC pretreatment exhibited good predictive power with no misclassification. This study may provide a novel perspective to employ a comprehensive analytical approach to brand discrimination of CCA. The synergetic strategy based on LIBS together with NIR offers atomic and molecular information of CCA, which could be exemplary for future research on the rapid discrimination of TCM.

Quality consistency evaluation on three species of North Patrininae herba by high-performance liquid chromatography coupled with electrospray ion trap time-of-flight mass spectrometry and network pharmacology approaches.

North Patrininae herba, a perennial herbaceous plant, has long been used in traditional Chinese medicine to treat appendicitis, enteritis, and dysentery. Sonchus arvensis L., Sonchus oleraceus L., and Ixeris chinensis (Thunb.) Nakai are used as substitutes for North Patrininae in different regions, but the consistency of chemical composition and efficacy of these three species is still unknown. In this study, a detailed chemical analysis was carried out of the extract obtained from Sonchus arvensis L., Ixeris chinensis (Thunb.) Nakai and Sonchus oleraceus L. and a chemical component not previously reported in Ixeris chinensis (Thunb.) Nakai was found-Luteolin-7-O-(6''-malonylglucoside). The mechanism of action of the extract against inflammation and type II diabetes was investigated using network pharmacology and analysis of blood-absorbed components following oral dosing of rats. Finally, a highly accurate and reliable method was established for quality control purposes. The results showed that Sonchus arvensis L. and Sonchus oleraceus L. may be considered potential resources of a medicinal compound, whereas Ixeris chinensis (Thunb.) Nakai requires further evaluation.

Peptide Biomarkers Discovery for Seven Species of Deer Antler Using LC-MS/MS and Label-Free Approach.

Deer antler is a globally widely used precious natural medicine and the material of deer horn gelatin. However, identification of deer antler species based on traditional approaches are problematic because of their similarity in appearance and physical-chemical properties. In this study, we performed a comprehensive antler peptidome analysis using a label-free approach: nano LC-Orbitrap MS was applied to discover peptide biomarkers in deer adult beta-globin (HBBA), and HPLC-Triple Quadrupole MS was used to verify their specificity. Nineteen peptide biomarkers were found, on which foundation a strategy for antlers and a strategy for antler mixtures such as flakes or powder are provided to identify seven species of deer antler including Eurasian elk (Alces alces), reindeer (Rangifer tarandus), white-tailed deer (Odocoileus viginianus), white-lipped deer (Przewalskium albirostris), fallow deer (Dama dama), sika deer (Cervus nippon), and red deer (Cervus elaphus) simultaneously. It is worth noting that our search found that the HBBA gene of sika deer, red deer, and North American wapiti (Cervus canadensis) in China may have undergone severe genetic drifts.

Management of imported malaria cases and healthcare institutions in central China, 2012-2017: application of decision tree analysis.

BACKGROUND: Imported malaria has been an important challenge for China. Fatality rates from malaria increased in China, particularly in Henan Province, primarily due to malpractice and misdiagnoses in healthcare institutions, and the level of imported malaria. This study aims to investigate the relationship between the state of diagnosis and subsequent complications among imported malaria cases at healthcare institutions, based on malaria surveillance data in Henan Province from 2012 to 2017. METHODS: A retrospective descriptive analysis was performed using data from the Centre for Disease Control and Prevention, Zhengzhou City, the capital of Henan Province. A decision tree method was exploited to provide valuable insight into the correlation between imported malaria cases and healthcare institutions. RESULTS: From 2012 to 2017, there were 371 imported malaria cases, mostly in males aged between 20 and 50 years, including 319 Plasmodium falciparum cases. First visits of 32.3%, 19.9% and 15.9% malaria cases for treatment were to provincial, municipal and county healthcare institutions, respectively. The time interval between onset and initial diagnosis of 284 cases (76.5%) and the time interval between initial diagnosis and final diagnosis of 197 cases (53.1%) was no more than 72 h. An apparent trend was found that there were notably fewer patients misdiagnosed at first visit to healthcare institutions of a higher administrative level; 12.5% of cases were misdiagnosed in provincial healthcare institutions compared to 98.2% in private clinics, leading to fewer complications at healthcare institutions of higher administrative level due to correct initial diagnosis. In the tree model, the rank of healthcare facilities for initial diagnosis, and number of days between onset and initial diagnosis, made a major contribution to the classification of initial diagnosis, which subsequently became the most significant factor influencing complications developed in the second tree model. The classification accuracy were 82.2 and 74.1%, respectively for the tree models of initial diagnosis and complications developed. CONCLUSION: Inadequate seeking medical care by imported malaria patients, and insufficient capacity to diagnose malaria by healthcare institutions of lower administrative level were identified as major factors influencing complications of imported malaria cases in Henan Province. The lack of connection between uncommon imported malaria cases and superior medical resources was found to be the crucial challenge. A web-based system combined with WeChat to target imported malaria cases was proposed to cope with the challenge.

Discovery and identification of interspecies peptide biomarkers in the seahorse species using liquid chromatography tandem mass spectrometry and chemometrics.

Seahorses have important edible and medicinal values including strengthening the body, tonifying the liver and kidneys, and reducing swelling. And there are abundant seahorse species on Earth. Many seahorses have large price differences due to the scarcity of resources, and some seahorses with similar appearances appear to be confused for use. While in market trading, Hippocampus is susceptible to loss of specialized morphology characteristics, making it difficult to distinguish between specific species. Here we report an effective method based on peptide biomarkers for the identification of seahorse species. Peptide biomarkers for each species were predicted using nano-liquid chromatography-tandem mass spectrometry (Nano-LC-MS/MS) combined with chemometrics software. One unique biomarker peptide for each species was synthesized and verified, and finally developed a liquid chromatography tandem mass spectrometry (LC-MS/MS) multiple reaction monitoring method. The results indicate that the method has great potential for species-specific identification of seahorses and their preparations, among others.

Discovery of species-specific peptide markers and development of quality-evaluation strategies for deer horn gelatin using liquid chromatography-tandem mass spectrometry and a label-free methodology.

Deer horn gelatin (DHG) is a valuable nutritional dietary supplement. Due to the significant variation in the price of DHG from different sources, it is important to assess its quality and to clarify the species of its raw material. However, due to the similarity in appearance and physicochemical properties, as well as the destruction of genetic material during the manufacturing process, it is difficult to distinguish DHG from gelatin derived from other sources. Furthermore, current methods are unable to evaluate the overall quality of DHG. Using Nano LC-Orbitrap MS and data analysis software, DHG samples from five deer species were analyzed to identify peptide markers specific to alpha-2-HS-glycoprotein (AHSG) and collagen. The peptide markers were validated using HPLC-Triple Quadrupole MS, and strategies for assessing the quality of DHG were developed. Eighteen peptide markers were discovered, comprising peptides with differing specificities. Three strategies for the identification, characteristic mapping, and content determination of DHG were developed. These strategies can be used to assess the quality of deer gelatin.

Bioactive sesquiterpenoids and diterpenoids from the liverwort Bazzania albifolia.

One undescribed 6,7-secofusicoccane-type diterpene, albifolione (1), and one new aromadendrane-type sesquiterpenoid ketone, methyl 2-oxoaromadendra-1(10),3-dien-12-oate (2), along with four known compounds, δ-cuparenol (3), fusicoauritone (4), chiloscyphenols A and B (5 and 6, resp.), were isolated from the liverwort Bazzania albifolia Horik. The structures and relative configurations of 1 and 2 were established unequivocally on the basis of spectroscopic data analysis. Preliminary bioassay revealed that compound 5 showed quite strong cytotoxicity against the human tumor cell line MCF-7, with an IC(50) value of 5.6 µM.

Quality Evaluation of Traditional Chinese Medicine Prescription in Naolingsu Capsule Based on Combinative Method of Fingerprint, Quantitative Determination, and Chemometrics.

Background: Naolingsu capsule (NLSC) is a well-known traditional Chinese medicine (TCM) prescription in China. It is widely used to treat neurasthenia, insomnia, cardiovascular and cerebrovascular disease, and other diseases. However, its inalienable chemical groups have not been carried out. Methods: We first established the nontargeted investigation based on fingerprinting coupled with UHPLC-Q/TOF-MS/MS. Second, the quantitative methods based on HPLC-DAD and LC-MS/MS were connected to the synchronous quantitative assurance of eleven and fourteen marker compounds. Finally, the quantitative information was processed with SIMCA-P for differentiating the distinctive bunches of samples to screen the foremost appropriate chemical markers. Results: The similarity of HPLC fingerprints of 24 batches of NLSC samples was 0.645-0.992. In total, 37 flavonoids, 21 organic acids, 22 lignans, 13 saponins, and 20 other compounds were recognized in NLSC by the UHPLC-Q/TOF-MS/MS method. The quantitative determination was approved for linearity, discovery limits, accuracy, repeatability, soundness, and precision. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) models accomplished the great classification of the samples from the five enterprises, respectively. Rehmannioside D (RD), methylophiopogonanone A (MPA), 3,6'-disinapoyl sucrose (DS), schisandrin B (SSB), epimedin C (EC), icariin (ICA), and jujuboside B (JB) were considered as the potential chemical markers for NLSC quality control. Conclusion: The experimental results illustrated that the combinative strategy was valuable for quick pharmaceutical quality assessment, which can potentially differentiate the origin, decide the realness, and assess the overall quality of the formulation.

Identification of chemosensitizing agents of colorectal cancer in Rauvolfia vomitoria using an NMR-based chemometric approach.

Searching for new adjuvants of conventional chemotherapeutic approaches against colorectal cancer cells is extremely urgent. In current research, a non-targeted analytical approach was established by combining proton nuclear magnetic resonance spectroscopy with a chemometrics data mining tool to identify chemosensitizing agents from Rauvolfia vomitoria. This approach enabled the identification of potential active constituents in the initial fractionation process and provided their structural information. This strategy was validated by its application to Rauvolfia vomitoria extract exhibiting chemosensitizing activity on 5-fluorouracil against colorectal cancer cells. After the workflow, the biochemometrics analysis showed that at least 15 signals (Variable influence on projection (VIP) > 1) could have contributions in the differentiation of various fractions. Through systematic literature and database searches, we found that the most active fraction (fraction 7) exhibited the highest presence of sabazin-type and armaniline-type alkaloids, which were potential chemosensitizers as previously reported. To validate the results of the strategy, the effect of 5-FU and compounds isolated from fraction seven incubation on HCT-8 and LoVo cell vialibilty were evaluated. These results evidenced that compound ß-carboline (3), 1-methyl-ß-carboline (4), and lochnerine (6) could enhance the cytotoxicity of 5-fluorouracil against to Colorectal cancer cells. Besides, 21 compounds including two new compounds were isolated from Rauvolfia vomitoria. The experimental results verify the reliability of the method, and this approach provides a new and efficient tool to overcome some of the bottlenecks in natural products drug discovery.

Phenolic compounds with NF-κB inhibitory effects from the fungus Phellinus baumii.

Chemical investigation of the fungus Phellinus baumii has resulted in characterization of five previously undescribed hispidin derivatives, phellibaumins A-E (1-5), as well as two pairs of new non-equivalent epimeric benzyl dihydroflavones, methylphelligrin A (9), epi-methylphelligrin A (10), methylphelligrin B (11), and epi-methylphelligrin B (12), together with five known compounds, interfungin B (6), phelligridin H (7), phelligridimer A (8), phelligrin A (13), and epi-phelligrin A (14). Phellibaumin A (1) was a novel hispidin derivative with a unique 3,4-dihydroxybenzofuran unit. These compounds exhibited NF-κB inhibitory activity with IC(50) values of 52.96 µM (1), 41.40 µM (2), 52.92 µM (5), 36.44 µM (9 and 10), and 22.46 µM (11 and 12), respectively.

TaMYB29: A Novel R2R3-MYB Transcription Factor Involved in Wheat Defense Against Stripe Rust.

Members of the R2R3-MYB transcription factor superfamily have been implicated in plant development, improved disease resistance, and defense responses to several types of stresses. To study the function of TaMYB29 transcription factor-a member of the R2R3-MYB superfamily-in response to an avirulent race of stripe rust pathogen, Puccinia striiformis f. sp. tritici (Pst), we identified and cloned the TaMYB29 gene from wheat cultivar (cv.) AvS+Yr10 following infection with Pst. The TaMYB29 protein, comprising 261 amino acids, contains two highly conserved MYB domains. We first showed that TaMYB29 is a transcription factor, whose transcriptional levels are significantly induced by salicylic acid (SA), abscisic acid (ABA), jasmonic acid (JA), ethylene (ET), and Pst. The results showed that TaMYB29 is involved in the wheat response to stipe rust. The overexpression of the TaMYB29 gene resulted in the accumulation of reactive oxygen species (ROS) and pathogen-independent cell death in Nicotiana benthamiana leaves. The silencing of TaMYB29 gene in wheat cv. AvS+Yr10, containing the stripe rust resistance gene Yr10, promoted hyphae growth, significantly downregulated the expression of pathogenesis-related (PR) genes, and substantially reduced the wheat resistance to Pst compared with the non-silenced control. In addition, the accumulation of hydrogen peroxide (H2O2) significantly decreased, and the activity of catalase, an enzyme required for H2O2 scavenging, was elevated. Altogether, TaMYB29 positively regulates the defense response against stripe rust in wheat AvS+Yr10 by enhancing H2O2 accumulation, PR gene expression, and SA signaling pathway-induced cell death. These results provide new insights into the contribution of TaMYB29 to the defense response against rust pathogens in wheat.

Spectroscopic methodologies and molecular docking studies on the interaction of antimalarial drug piperaquine and its metabolites with human serum albumin.

Artemisinin-based combination therapy is widely used for the treatment of uncomplicated Plasmodium falciparum malaria, and piperaquine (PQ) is one of the important partner drugs. During the biotransformation of PQ, M1 (N-oxidation product), M2 (N-oxidation product), M3 (carboxylic acid product), M4 (N-dealkylation product), and M5 (N-oxidated product of M4) are formed by cytochrome P450 pathways. Despite decades of clinical use, the interactions between PQ and its main metabolites (PQs) with human serum albumin (HSA) have not been reported. In the present study, the binding of PQs with HSA under physiological conditions was investigated systematically through fluorescence, circular dichroism (CD) spectroscopy, and molecular docking methods. The experimental results show that the intrinsic fluorescence quenching of HSA was induced by those compounds resulting from the formation of stable HSA-compound complexes. The main forces involved in the interactions between PQ, M1, and M2 which bind to HSA were hydrogen s and van der Waals forces, while the interactions of M3, M4, and M5 were driven by hydrophobic forces. The main binding sites of the compounds to HSA were also examined by classical fluorescent marker experiments and molecular docking studies. Binding constants (Kb) revealed that the affinities of the PQ, M1, M2, M3, and M4 to HSA were stronger than that of M5. Additionally, the binding rates of PQs with HSA were determined by ultrafiltration methods. Consistent with the binding constant results, the binding rate of M5 was lower than the binding rates of PQ, M1, M2, M3, and M4. Furthermore, PQs binding to HSA led to conformational and structural alterations of HSA, as revealed by multi-spectroscopic studies. In order to investigate one possible mechanism by which PQs inhibit the growth of malaria-causing Plasmodium parasites, 1H NMR spectroscopy was performed to investigate the interaction of the PQs with heme. This study is beneficial to enhance our understanding of the ecotoxicology and environmental behaviors of PQ and its metabolites.

Spectroscopic methodologies and molecular docking studies on the interaction of the soluble guanylate cyclase stimulator riociguat with human serum albumin.

Interaction of riociguat with human serum albumin (HSA) is extremely important in understanding the drug's disposition and efficiency. In the current study, the binding of riociguat to HSA was explored using spectroscopic methods and molecular docking. The quenching constant, the binding constant, the number of binding sites, thermodynamic parameters, and the secondary structure of protein were determined. A fluorescence study revealed that riociguat quenched HSA fluorescence via static quenching with a binding constant of 1.55 × 104 L mol-1 at 298 K. The calculated thermodynamic parameters indicated that the binding process was spontaneous and that the main interaction force was hydrophobic interaction. Site marker competitive binding experiments and molecular docking studies suggested that riociguat was inserted into the subdomain IIA (site I) of HSA. Alterations in the protein secondary structure after drug complexation were predicted. Results indicated that the protein a-helix structure increased with an increasing concentration of riociguat. This indicated that a riociguat-HSA complex was formed and that the protein secondary structure was altered by the addition of riociguat.

Aromatic compounds from the liverwort Conocephalum japonicum.

Two undescribed dimeric ArC2 derivatives, cis- and trans-1,2-bis(3,4-dimethoxyphenyl)cyclobutane (1 and 2), one new monoterpenes esters, 2alpha,5beta-dihydroxybornane-2-cis-cinnamate (3), along with eight known compounds, 2alpha,5beta-dihydroxybornane-2-trans-cinnamate (4), perrottetin E (5), isoriccardin C (6), marchantin A (7), marchantin E (8), marchantin C (9), and isomarchantin C (10) were isolated from the liverwort Conocephalum japonicum. All the structures were established by extensive spectroscopic analysis. The isolated compounds 3-10 were evaluated for their cytotoxicity against the human KB cell line with IC50 values ranging from 16.5 to 50.2 microM.

Scaparvin A, a novel caged cis-clerodane with an unprecedented C-6/C-11 bond, and related diterpenoids from the liverwort Scapania parva.

A novel caged cis-clerodane diterpenoid, scaparvin A, possessing an unprecedented C-6/C-11 bond and a ketal ring, as well as four new cis-clerodane derivatives, scaparvins B-E, were isolated from the Chinese liverwort Scapania parva. Their absolute structures were elucidated by analysis of NMR and CD data coupled with electronic circular dichroism (ECD) calculations. It was proposed that an enzymatic intramolecular aldol reaction was the key step in the biogenetic pathway of scaparvin A.

Labdane diterpenoids and highly methoxylated bibenzyls from the liverwort Frullania inouei.

Four undescribed labdane diterpenoids, 1,2-dehydro-3,7-dioxo-manoyl oxide (1), 1,2-dehydro-7 beta-hydroxy-3-oxo-manoyl oxide (2), 3,7-dioxo-manoyl oxide (3), and 3beta-hydroxy-7-oxo-manoyl oxide (4) together with three known diterpenoids (5-7) and four highly methoxylated bibenzyls (8-11) were isolated from the liverwort Frullania inouei. The absolute structures of 1-4 were established by combined analysis of NMR data, CD data coupled with TDDFT CD calculations, and single-crystal X-ray diffraction measurement. Cytotoxicity tests to human tumor KB, KB/VCR, K562 or K562/A02 cells showed bibenzyls 8-11 inhibited cell proliferation with ID(50) values ranging from 11.3 to 49.6 microM and overcame the multidrug resistance (MDR) with the reversal fold (RF) values ranging from 3.19 to 10.91 (5 microM) for vincristine-resistant KB/VCR and RF values from 4.40 to 8.26 (5 microM) for adriamycin-resistant K562/A02 cells, respectively. However, none of the diterpenoids were found to be active (ID(50)>50 microM).

Secondary metabolites from the liverwort Ptilidium pulcherrimum.

A new trinortriterpenoid, diospyrolide acetate (1) and a new diphenylmethane derivative, pulcherrimumin (12), together with ten known pentacyclic triterpenoids (2-11) and four aromatic compounds (13-16), were isolated from the liverwort Ptilidium pulcherrimum. Their structures were established on the basis of extensive analysis of NMR data and by chemical methods. The cytotoxicity of compounds 1-16 was evaluated against the PC3, MDA-MB-231, and Hela cells lines. Ursane triterpenoids 8-10 exhibited moderate cytotoxicity against PC3 cells with IC50 values ranging from 10.1 to 39.7 microM.