Rapid characterization of non-volatile phenolic compounds reveals the reliable chemical markers for authentication of traditional Chinese medicine Xiang-ru among confusing Elsholtzia species.

The aerial parts of Mosla chinensis Maxim. and Mosla chinensis cv. 'Jiangxiangru' (MCJ) are widely utilized in traditional Chinese medicine (TCM), known collectively as Xiang-ru. However, due to clinical effectiveness concerns and frequent misidentification, the original plants have increasingly been substituted by various species within the genera Elsholtzia and Mosla. The challenge in distinguishing between these genera arises from their similar morphological and metabolic profiles. To address this issue, our study introduced a rapid method for metabolic characterization, employing high-resolution mass spectrometry-based metabolomics. Through detailed biosynthetic and chemometric analyses, we pinpointed five phenolic compounds-salviaflaside, cynaroside, scutellarein-7-O-D-glucoside, rutin, and vicenin-2-among 203 identified compounds, as reliable chemical markers for distinguishing Xiang-ru from closely related Elsholtzia species. This methodology holds promise for broad application in the analysis of plant aerial parts, especially in verifying the authenticity of aromatic traditional medicinal plants. Our findings underscore the importance of non-volatile compounds as dependable chemical markers in the authentication process of aromatic traditional medicinal plants.

Stepwise changes in flavonoids in spores/pollen contributed to terrestrial adaptation of plants.

Protecting haploid pollen and spores against UV-B light and high temperature, 2 major stresses inherent to the terrestrial environment, is critical for plant reproduction and dispersal. Here, we show flavonoids play an indispensable role in this process. First, we identified the flavanone naringenin, which serves to defend against UV-B damage, in the sporopollenin wall of all vascular plants tested. Second, we found that flavonols are present in the spore/pollen protoplasm of all euphyllophyte plants tested and that these flavonols scavenge reactive oxygen species to protect against environmental stresses, particularly heat. Genetic and biochemical analyses showed that these flavonoids are sequentially synthesized in both the tapetum and microspores during pollen ontogeny in Arabidopsis (Arabidopsis thaliana). We show that stepwise increases in the complexity of flavonoids in spores/pollen during plant evolution mirror their progressive adaptation to terrestrial environments. The close relationship between flavonoid complexity and phylogeny and its strong association with pollen survival phenotypes suggest that flavonoids played a central role in the progression of plants from aquatic environments into progressively dry land habitats.

Anti-senescence effects of Rhodiola crenulate extracts on LO2 cells and bioactive compounds.

ETHNOPHARMACOLOGICAL RELEVANCE: Rhodiola crenulata (Rc) is a traditional herb, used in Tibetan medicine, has shown promise efficacy in physical performance improvement, work capacity enhancement, fatigue elimination, and altitude sickness prevention. Also, Rc exhibited therapeutic effects on aging-related diseases. However, relevant researches on Rc and their bioactive components are quite few and needs further investigation. AIM OF THE STUDY: The objective of this study was to understand the relationship between phytochemical profiles and their activities of Rc extracts. MATERIALS AND METHODS: Rc extracts prepared by solvents with various hydrophilicity (i.e. aqueous ethanol (70%, v/v), water, and ethyl acetate), and their chemical compositions and specific compounds were analyzed by chemical analysis method and ultra-performance liquid chromatography quadruple time-of-flight mass spectrometry (UPLC-QTOF-MS). The regulate effects of Rc extracts on senescence and antioxidant activity were evaluated using the models of LO2 cells and Caenorhabditis elegans. RESULTS: The 70% ethanol extracts exhibited better regulating effects on senescence via the assays of senescence -associated ß-galactosidase (SAßG) staining and lifespan, which was consistent with the higher antioxidant activities observed based on the results of antioxidant assays. A total of 14 phytochemicals have been identified in 70% ethanol extracts, whereas the other two extracts contained much fewer compounds in varieties. Phytochemical profile of water extract was similar to the first half (polar compounds, running time: 0-6 min) of 70% ethanol extract profile, while those of ethyl acetate extract was consistent with its second half (more nonpolar compounds, running time: 6-12 min). CONCLUSIONS: The 14 phytochemicals in Rc might exhibit additive or synergistic effects on senescence regulating and antioxidant activities, providing theoretical basis for daily administration of Rc.

Metabolic profiling integrated with pharmacokinetics to reveal the material basis of Xiaokeyinshui extract combination in the treatment of type 2 diabetes in rats.

Xiaokeyinshui extract combination (XEC), originating from a traditional Chinese formula Xiaokeyinshui (XKYS) recorded in ancient Bencao, has been reported to exert significant hypoglycemic effects. However, the chemical profiles, metabolic transformation and pharmacokinetic behavior of XEC in vivo were unclear. The research was to investigate the chemical constituents, metabolic profiles and pharmacokinetic behavior of XEC. A UPLC-QE-Orbitrap-HRMS qualification method was developed to identify the chemical constituents in XEC and xenobiotics of XEC in plasma, urine, feces and bile of rats after oral administration. A LC-MS quantification method was established and applied for the pharmacokinetic studies of major active compounds of XEC in normal and T2DM rats and Coptidis Rhizoma extracts (CRE) in T2DM rats. Fifty eight compounds in XEC and a total of 152 xenobiotics were identified in T2DM rats, including 28 prototypes and 124 metabolites. The metabolic pathways were demethylation, demethyleneization, reduction, hydroxylation, hydrolysis and subsequent binding reactions, including glucuronidation, sulfation and methylation. According to the results of chemical constituents and metabolites, 7 ingredients, including berberine, palmatine, coptisine, epiberberine, berberrubine, magnoflorine and aurantio-obtusin were suggested for markers to comparative pharmacokinetics study in normal rats and T2DM rats. Compared with normal rats, the Tmax of berberine, palmatine, coptisine, epiberberine, berberrubine and magnoflorine was significantly longer. The value of Cmax for palmatine, coptisine, epiberberine and berberrubine was significantly decreased in XEC T2DM group. The value of AUC for alkaloids was higher in diabetic rats. After oral CRE, alkaloids including berberine, palmatine, coptisine, epiberberine, berberrubine and magnoflorine could be detected in vivo. Compared with T2DM rats after oral administration of CRE, the value of Tmax and Cmax for berberine, palmatine, coptisine, epiberberine, berberrubine and magnoflorine exhibited significant differences in XEC T2DM group. This research provided an overview of the chemical profiles and metabolic profiling of XEC and elucidated the effect of diabetic state and compatibility on pharmacokinetic behaviors of active components in XEC. This research also can provide the material basis of XEC for subsequent quality control research.

Optimization of extraction process of Chuanxiong Rhizoma-Gastrodiae Rhizoma based on network pharmacology and multi-index orthogonal test / 中国中药杂志

To optimize the extraction process of Chuanxiong Rhizoma-Gastrodiae Rhizoma herb pair by network pharmacology combined with analytic hierarchy process(AHP)-entropy weight method and multi-index orthogonal test. The potential active components and targets of Chuanxiong Rhizoma-Gastrodiae Rhizoma were screened by network pharmacology and molecular docking, and the process evaluation indexes were determined with reference to the Chinese Pharmacopoeia(2020 edition). The core components of Chuanxiong Rhizoma-Gastrodiae Rhizoma were determined as gastrodin, parishin B, parishin C, parishin E, ferulic acid, and 3-butylphthalide. With the extraction volume of each indicator and yield of dry extract as comprehensive evaluation indicators, the extraction conditions were optimized by the AHP-entropy weight method and orthogonal test as the ethanol volume of 50%, the solid-liquid ratio of 1∶8(g·mL~(-1)), extraction for three times, and 1.5 h each time. Through network pharmacology and molecular docking, the process evaluation index was determined, and the optimized process was stable and reproducible for the extraction of Chuanxiong Rhizoma-Gastrodiae Rhizoma herb pair, which could provide reference for in-depth research.

Huoxin Pill Reduces Myocardial Ischemia Reperfusion Injury in Rats via TLR4/NFκB/NLRP3 Signaling Pathway / 中国结合医学杂志

OBJECTIVE@#To explore the protective effect of Huoxin Pill (HXP) on acute myocardial ischemia-reperfusion (MIRI) injury in rats.@*METHODS@#Seventy-five adult SD rats were divided into the sham-operated group, model group, positive drug group (diltiazem hydrochloride, DH), high dose group (24 mg/kg, HXP-H) and low dose group (12 mg/kg, HXP-L) of Huoxin Pill (n=15 for every group) according to the complete randomization method. After 1 week of intragastric administration, the left anterior descending coronary artery of the rat's heart was ligated for 45 min and reperfused for 3 h. Serum was separated and the levels of creatine kinase (CK), creatine kinase isoenzyme (CK-MB) and lactate dehydrogenase (LDH), superoxide dismutase (SOD), and malondialdehyde (MDA), hypersensitive C-reactive protein (hs-CRP) and interleukin-1β (IL-1β) were measured. Myocardial ischemia rate, myocardial infarction rate and myocardial no-reflow rate were determined by staining with Evans blue and 2,3,5-triphenyltetrazolium chloride (TTC). Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine (BATMAN) databases were used to screen for possible active compounds of HXP and their potential therapeutic targets; the results of anti-inflammatory genes associated with MIRI were obtained from GeneCards, Drugbank, Online Mendelian Inheritance in Man (OMIM), and Therapeutic Target Datebase (TTD) databases was performed; Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment were used to analyze the intersected targets; molecular docking was performed using AutoDock Tools. Western blot was used to detect the protein expression of Toll-like receptor 4 (TLR4)/nuclear factor kappa-B (NFκB)/NOD-like receptor protein 3 (NLRP3).@*RESULTS@#Compared with the model group, all doses of HXP significantly reduced the levels of LDH, CK and CK-MB (P<0.05, P<0.01); HXP significantly increased serum activity of SOD (P<0.05, P<0.01); all doses of HXP significantly reduced the levels of hs-CRP and IL-1β (P<0.05, P<0.01) and the myocardial infarction rate and myocardial no-reflow rate (P<0.01). GO enrichment analysis mainly involved positive regulation of gene expression, extracellular space and identical protein binding, KEGG pathway enrichment mainly involved PI3K-Akt signaling pathway and lipid and atherosclerosis. Molecular docking results showed that kaempferol and luteolin had a better affinity with TLR4, NFκB and NLRP3 molecules. The protein expressions of TLR4, NFκB and NLRP3 were reduced in the HXP group (P<0.01).@*CONCLUSIONS@#HXP has a significant protective effect on myocardial ischemia-reperfusion injury in rats, and its effect may be related to the inhibition of redox response and reduction of the inflammatory response by inhibiting the TLR4NFκB/NLRP3 signaling pathway.

Different acupotomy for stenosing tenosynovitis: A protocol for systematic review and network meta-analysis.

BACKGROUND: Stenosing tenosynovitis (STS) is a chronic aseptic inflammation caused by mechanical friction. The main clinical manifestations are local pain and limited activity of the affected parts, which reduce people's quality of life. The clinical effect of acupotomy in the treatment of STS is significant, and the operation is simple and the side effect is small. But there are many kinds of acupotomology, and there is a lack of comparative study between different Acupotomology. In this study, the effectiveness of 4 commonly used needle knife therapies (v-knife, oblique knife, crochet knife, flat knife) was ranked by the method of network meta. METHODS: CNKI, Wanfang, VIP, Sinomed, PubMed, and Cochrane Library were searched to collect randomized controlled trials of v-knife, oblique knife, crochet knife, and flat knife in the treatment of STS. The search time limit is from the date of establishment to October 15, 2021. Revman5.3, gemtc 0.14.3, and stata14.2 were used for data analysis, and Cochrane bias risk assessment tool was used to screen and evaluate the quality of included literatures. CONCLUSION: Objective to provide evidence-based medicine evidence for clinical selection of the best needle knife treatment scheme for STS.

Biosynthesis-based spatial metabolome of Salvia miltiorrhiza Bunge by combining metabolomics approaches with mass spectrometry-imaging.

Defining the spatial distributions of metabolites and their structures are the two key aspects for interpreting the complexities of biosynthesis pathways in plants. As a means of obtaining information on the spatial distribution of metabolites, a strategy is needed that has high sensitivity and allows visualization. Toward this goal, we carried an untargeted metabolomics to obtain detailed metabolic information on different plant parts of Salvia miltiorrhiza, the roots of which are widely used in traditional Chinese medicine. Systematic optimization of desorption electrospray ionization mass spectrometry imaging (DESI-MSI) including parameter selection and sample preparation were carried out to improve the sensitivity of the method for plant samples. Guided by the metabolomics data, the spatial distributions of diverse metabolites, including phenolic acids, flavonoids, tanshinones, carbohydrates, and lipids, were characterized and visualized for both the underground and aerial parts. To integrate the information pertaining to the spatial distribution of metabolites, the flavonoids and phenolic acids (phenylpropanoid metabolic pathway) were chosen as examples for in-depth study the biosynthesis pathways in S. miltiorrhiza. The complementary data obtained from the metabolomics study and mass spectrometry imaging enabled the identification of key reactions involved in flavonoid biosynthesis in flowers, which lead the changes in metabolite distribution. The analysis also identified the core precursor for phenolic acid biosynthesis in Salvia species. Therefore, the powerful combination of metabolomics and mass spectrometry imaging provides a basis for obtaining detailed information on spatial metabolome and constitutes a platform for deep understanding the biosynthesis of bioactive metabolites in plants.

Structural elucidation of lignin macromolecule from abaca during alkaline hydrogen peroxide delignification.

To maximize the utilization of Abaca lignin in the current biorefinery, structural characteristics of native lignin from Abaca were firstly comprehensively investigated. Parallelly, effective delignification of Abaca was achieved by alkaline hydrogen peroxide (AHP) process, which facilitated the production of specialty paper in industry. The structural changes of lignin macromolecules during the AHP delignification were illustrated by comparing the structural differences of the released lignin fraction and corresponding native lignin, which were analyzed via the advanced analytical methods, such as 2D-HSQC NMR, 31P NMR, pyrolysis-GC/MS, and GPC techniques. It was found that Abaca lignin is a HGS-type lignin, which is overwhelmingly composed of ß-O-4 linkages and abundant hydroxycinnamic acids (mainly p-coumaric acid). In addition, partial cleavage of ß-O-4 linkages and p-coumarate in lignin occurred during the AHP delignification process. Meanwhile, AHP process also led to the elevation of H-type lignin units in AHPL. Considering that ß-O-4 bond is vulnerable in the catalytic degradation process of lignin, the lignin with abundant ß-O-4 linkages is beneficial to the downstream conversion of lignin into aromatic chemicals.

Structural Features of Alkaline Dioxane Lignin and Residual Lignin from Eucalyptus grandis × E. urophylla.

In the present study, lignin from eucalyptus was extracted with 80% alkaline dioxane (0.05 M NaOH) from ball-milled wood and subsequently fractionated by gradient acid precipitation from the filtrate. Meanwhile, the residual lignin was prepared by a double enzymatic hydrolysis process. The yield of the lignin extracted by alkaline dioxane (LA-2) was 29.5%. The carbohydrate contents and molecular weights of the gradient acid precipitated lignin fractions gradually decreased from 4.90 to 1.36% and from 7770 to 5510 g/mol, respectively, with the decline of the pH value from 6 to 2. Results from two-dimensional heteronuclear single quantum coherence nuclear magnetic resonance (NMR) and 31P NMR spectroscopy showed an evident reduction of ß- O-4 ' linkages with the pH value decrease, while the contents of aliphatic -OH, phenolic -OH, and carboxylic groups displayed an increasing trend. Moreover, the residual lignin exhibited the highest molecular weight (11690 g/mol), the most abundant ß- O-4 ' linkages (71.1%), and the highest S/G ratio (4.68).

[Effect of ningxintong granule in treating patients of coronary heart disease with left ventricular diastolic dysfunction and qi-deficiency blood-stasis syndrome].

OBJECTIVE: To investigate the effect of Ningxintong Granule (NXTG) on left ventricular diastolic dysfunction (LVDD) in patients with coronary heart disease (CHD) of qi-deficiency and blood-stasis syndrome in Chinese medicine. METHODS: Seventy-six CHD patients with LVDD of qi-deficiency blood-stasis syndrome were randomized into two groups. The 36 patients in the control group were treated by Western medical conventional treatment alone, and 40 patients in the treated group were treated by NXTG (Consisting of milk vetch root, pueraria root, ligustici, ilex-puhesceus, manchurian wildginger, 10 g in each package) and Western medical conventional treatment in combination, one package each time, thrice daily, 4 weeks as one therapeutic course. The left ventricular function (LVF) of all patients (including Emas, Amas, E/A, DT and IVRT) was evaluated by Doppler echocardiography before and after treatment, and the changes of clinical effectiveness and TCM syndrome effectiveness were observed. RESULTS: Compared with the control group after treatment, Emas, E/A increased, while Amas, DT, IVRT decreased showing statistical difference (P < 0.05). The therapeutic effects in the treated group were significantly better (P < 0.05). CONCLUSION: NXTG with qi-enriching and blood-activating actions shows favorable effect in treating CHD patients with LVDD.

Effect of ningxintong granule in treating patients of coronary heart disease with left ventricular diastolic dysfunction and qi-deficiency blood-stasis syndrome / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of Ningxintong Granule (NXTG) on left ventricular diastolic dysfunction (LVDD) in patients with coronary heart disease (CHD) of qi-deficiency and blood-stasis syndrome in Chinese medicine.</p><p><b>METHODS</b>Seventy-six CHD patients with LVDD of qi-deficiency blood-stasis syndrome were randomized into two groups. The 36 patients in the control group were treated by Western medical conventional treatment alone, and 40 patients in the treated group were treated by NXTG (Consisting of milk vetch root, pueraria root, ligustici, ilex-puhesceus, manchurian wildginger, 10 g in each package) and Western medical conventional treatment in combination, one package each time, thrice daily, 4 weeks as one therapeutic course. The left ventricular function (LVF) of all patients (including Emas, Amas, E/A, DT and IVRT) was evaluated by Doppler echocardiography before and after treatment, and the changes of clinical effectiveness and TCM syndrome effectiveness were observed.</p><p><b>RESULTS</b>Compared with the control group after treatment, Emas, E/A increased, while Amas, DT, IVRT decreased showing statistical difference (P < 0.05). The therapeutic effects in the treated group were significantly better (P < 0.05).</p><p><b>CONCLUSION</b>NXTG with qi-enriching and blood-activating actions shows favorable effect in treating CHD patients with LVDD.</p>