Tea Polyphenols Reduce Inflammation of Orbital Fibroblasts in Graves' Ophthalmopathy via the NF-κB/NLRP3 Pathway.

OBJECTIVE: This study aimed to explore the effects of tea polyphenols (TP) on inflammation of orbital fibroblasts in Graves' ophthalmopathy (GO) and to provide new ideas for GO treatment. METHODS: Primary orbital fibroblasts were extracted from orbital adipose/connective tissues of patients with and without GO. Real-time quantitative PCR (RT-qPCR) was used to detect the expression of interleukin (IL)-6, IL-1ß, and monocyte chemotactic protein (MCP)-1 in non-GO and GO orbital fibroblasts. The CCK-8 assay was used to determine the appropriate concentration of TP for subsequent experiments. RT-qPCR and enzyme-linked immunosorbent assay (ELISA) were performed to investigate the effects of TP on lipopolysaccharide (LPS)-induced production of inflammatory cytokines. Nuclear factor-κB (NF-κB) expression was measured using Western blotting analysis. NOD-like receptor 3 (NLRP3) expression was detected using both Western blotting analysis and immunofluorescence staining. RESULTS: The mRNA levels of IL-6, IL-1ß, and MCP-1 in GO orbital fibroblasts were significantly higher than those in non-GO cells. TP treatment significantly inhibited LPS-induced production of inflammatory factors, including IL-6, IL-1ß, and MCP-1. TP also inhibited the expression levels of NF-κB and NLRP3. Inflammation in the GO orbital fibroblasts was higher than that in non-GO cells. TP inhibited the production of inflammatory cytokines in GO orbital fibroblasts in vitro through the NF-κB/NLRP3 pathway. CONCLUSION: These findings suggest that TP may have a potential role in GO treatment.

Meditation Using a Mobile App Improves Surgery Trainee Performance: A Simulation-Based Randomized Controlled Trial.

PURPOSE: To primarily investigate: (1) whether a 10-minute instant meditation practice using a mobile app could enhance arthroscopy performance and (2) whether a 10-day app-based meditation could reduce short-term arthroscopic skills deterioration. METHODS: Orthopaedic residents with no previous experience in arthroscopy and meditation were randomly assigned to groups A, B, and C. After initial standard competency-based arthroscopy training on the simulator on day 1, a pretest was performed via the simulator by all participants to assess their initial level of performance, then groups A and B were required to practice app-based mindfulness meditation 10 min/day for 10 consecutive days while group C did nothing. On day 11, all participants returned to perform a posttest. Before the posttest, the participants in group A practiced app-based meditation (10 minutes), whereas groups B and C had no intervention. RESULTS: In total, 43 participants were included and reached similar level of performance after initial training phase in day 1. On day 11, participants in group A had statistically a better instant arthroscopy performance than group B, with greater total score (mean difference [MD] 3.57; P < .001), less completion time (MD -42.89 seconds; P = .001), shorter camera (MD -23.38 cm; P < .001) and grasper (MD -15.23 cm; P = .002) path length, and less cartilage injury (MD -1.07%; P = .012). Participants in group B had less skills deterioration than group C, with better total score (MD -5.42; P < .001), less completion time (MD 51.96s; P = .002), camera path length (MD 28.41 cm; P = .007), and cartilage injury (MD 1.19%; P = .038). CONCLUSIONS: Meditation training using a mobile app enhanced instant simulation-based arthroscopy performance and reduced short-term skills deterioration of orthopaedic residents with no arthroscopy hands-on experience. CLINICAL RELEVANCE: A meditation using mobile app for clinicians and educators should be incorporated into simulation-based arthroscopy curriculums and perhaps clinical settings to improve arthroscopy performance and mental health of orthopaedic residents without any previous arthroscopy experience.

Modification of lysine deacetylation regulates curcumol-induced necroptosis through autophagy in hepatic stellate cells.

The excessive deposition of extracellular matrix (ECM) is the main characteristic of liver fibrosis, and hepatic stellate cells (HSCs) are the main source of ECM. The removal of activated HSCs has a reversal effect on liver fibrosis. Western blot and MTT analysis indicated that curcumol could relieve hepatic fibrosis by promoting HSCs receptor-interacting protein kinase 1/3 (RIP1/RIP3)-dependent necroptosis. Importantly, autophagy flow was monitored by constructing the mRFP-GFP-LC3 plasmid, and it was found that curcumol cleared activated HSCs in a necroptosis manner that was dependent on autophagy. Our study suggested that the activation of necrosome formed by RIP1 and RIP3 depended on Atg5, and that autophagosomes were also necessary for curcumol-induced necroptosis. Furthermore, microscale thermophoresis and co-immunoprecipitation assay results proved that curcumol could target Sirt1 to regulate autophagy by reducing the acetylation level of Atg5. The HSCs-specific silencing of Sirt1 exacerbated CCl4 -induced liver fibrosis in mice. The deacetylation of Atg5 not only accelerated the accumulation of autophagosomes but also enhanced the interaction between Atg5 and RIP1/RIP3 to induce necroptosis. Overall, our study indicated that curcumol could activate Sirt1 to promote Atg5 deacetylation and enhanced its protein-protein interaction function, thereby inducing autophagy and promoting the necroptosis of HSCs to reduce liver fibrosis.

Bioactivity-guided isolation of compounds from Sophora flavescens with antibacterial activity against Acinetobacter baumannii.

Bioactivity-guided fraction of an extract of Sophora flavescens to identify antibacterial compounds against Acinetobacter baumannii, led to the isolation of two new compounds, (2″R)-5-methoxy-7-hydroxy-8-lavandulylchromone (13) and (2S,ßS)-(-)-sophobiflavonoid CE (19), and 18 known flavonoids, (6aR,11aR)-(-)-maackiain (1), (2S)-(-)-8-prenylnaringenin (2), (2S)-(-)-exiguaflavanone K (3), (2S)-(-)-sophoraflavanone G (4), (2S)-(-)-leachianone A (5), (2S)-(-)-kushenol E (6), (2S)-(-)-leachianone G (7), (±)-kushenol F (8), (2S)-(-)-kurarinone (9), (2S)-(-)-kurarinol (10), (2 R,3R)- (+)-3,7,4'-trihydroxy-5-methoxy-8-prenylflavanone (11), (2S)-(-)-isoxanthohumol (12), (2S)-(-)-2'-methoxykurarinone (14), (2 R,3R)-(+)-kushenol I (15), calycosin (16), kuraridin (17), (2S)-(-)-kushenol A (18), and trifolirhizin (20). Their structures were elucidated based on NMR, MS, and CD spectroscopic analysis. Among them, 1, 2, 5, and 15 exerted modest antibacterial activity against A. baumannii, with MIC95 of 128-256 µg/mL for 2 and 256-512 µg/mL for 1, 5 and 15.

Association between neutrophils and renal impairment of rheumatoid arthritis: A retrospective cross-sectional study.

OBJECTIVE: Previous studies have shown that increased neutrophils, as a manifestation of oxidative stress, may be involved in the progression of kidney disease. To our knowledge, little is known about the relationship between neutrophils and renal impairment in rheumatoid arthritis (RA). Therefore, we aim to investigate whether neutrophil is associated with renal impairment in RA patients. METHODS: We retrospectively investigated the renal function of 602 RA patients in the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine by estimated glomerular filtration rate (eGFR) from September 2018 and September 2019. The exposure variable was neutrophils, and the main outcome was eGFR. General data (gender, age, duration, hypertension, diabetes, hobbies, and medication history), whole blood markers, lipid indexes, and inflammatory indexes were collected as much as possible. We used multivariable logistic regression analysis to evaluate the association between neutrophils and renal impairment in RA participants. RESULTS: A total of 89 cases (14.8%) had renal impairment with eGFR < 60 ml/min/1.73 m2 , and 75 cases (84.3%) were female. Subgroup analysis showed that female (odds ratio [OR] = 0.523, 95% confidence interval [CI]: 0.318-0.867, p = .011), neutrophils greater thsn 7.5 × 109 /L (OR = 2.314, 95% CI: 1.310-4.087, p = .004), NLR > 3.53 (OR = 1.757, 95% CI: 1.104-2.799, p = .018), hemoglobin less than 120 g/L (OR = 2.413, 95% CI: 1.418-4.118, p = .001), and UA > 360 µmol/L (OR = 6.052, 95% CI: 3.708-9.878, p < .001) was related to renal damage in RA. Adjusted for several confounders, the multivariable analysis indicated that neutrophils greater than 7.5 × 109 /L (OR = 1.784, 95% CI: 1.164-3.288, p = .031) was independently associated with an increased risk of renal impairment in RA. CONCLUSION: Our study demonstrated that neutrophils greater than 7.5 × 109 /L was associated with a high risk of renal impairment in RA, suggesting that neutrophil may be a biomarker for renal impairment in RA.

Recovery and recycling of solvent of counter-current chromatography: The sample of isolation of zeaxanthin in the Lycium barbarum L. fruits.

An efficient method of recovering and recycling solvent for counter-current chromatography was established by which zeaxanthin was separated from Lycium barbarum L. fruits. A column with activated carbon combined with high performance counter-current chromatography formed the recovering and recycling solvent system. Using the solvent system of n-hexane-ethyl acetate-ethanol-water (8:2:7:3, v/v) from the references, five injections were performed with an almost unchanged purity of zeaxanthin (80.9, 81.2, 81.5, 81.3, and 80.2% respectively) in counter-current chromatography separation. Meanwhile, the mobile phase reduced by half than conventional counter-current chromatography. By this present method, an effective improvement of counter-current chromatography solvent utilization was achieved.

Curcumol attenuates liver sinusoidal endothelial cell angiogenesis via regulating Glis-PROX1-HIF-1α in liver fibrosis.

OBJECTIVE: Hepatic sinusoidal angiogenesis owing to dysfunctional liver sinusoidal endothelial cells (LSECs) accompanied by an abnormal angioarchitecture is a symbol related to liver fibrogenesis, which indicates a potential target for therapeutic interventions. However, there are few researches connecting angiogenesis with liver fibrosis, and the deeper mechanism remains to be explored. MATERIALS AND METHODS: Cell angiogenesis and angiogenic protein were examined in primary LSECs of rats, and multifarious cellular and molecular assays revealed the efficiency of curcumol intervention in fibrotic mice. RESULTS: We found that curcumol inhibited angiogenic properties through regulating their upstream mediator hypoxia-inducible factor-1α (HIF-1α). The transcription activation of HIF-1α was regulated by hedgehog signalling on the one hand, and the protein stabilization of HIF-1α was under the control of Prospero-related homeobox 1 (PROX1) on the other. A deubiquitinase called USP19 could be recruited by PROX1 and involved in ubiquitin-dependent degradation of HIF-1α. Furthermore, our researches revealed that hedgehog signalling participated in the activation of PROX1 transcription probably in vitro. Besides, curcumol was found to ameliorate liver fibrosis and sinusoid angiogenesis via hedgehog pathway in carbon tetrachloride (CCl4 ) induced liver fibrotic mice. The protein expression of key regulatory factors, PROX1 and HIF-1α, was consistent with the Smo, the marker protein of Hh signalling pathway. CONCLUSIONS: In this article, we evidenced that curcumol controlling LSEC-mediated angiogenesis could be a promising therapeutic approach for liver fibrosis.

Gypenosides Prevent H2O2-Induced Retinal Ganglion Cell Apoptosis by Concurrently Suppressing the Neuronal Oxidative Stress and Inflammatory Response.

Our previous study demonstrated that gypenosides (Gp) exert protective effects on retinal nerve fibers and axons in a mouse model of experimental autoimmune optic neuritis. However, the therapeutic mechanisms remain unclear. Thus, in this study, a model of oxidative damage in retinal ganglion cells (RGCs) was established to investigate the protective effect of Gp, and its possible influence on oxidative stress in RGCs. Treatment of cells with H2O2 induced RGC injury owing to the generation of intracellular reactive oxygen species (ROS). In addition, the activities of antioxidative enzymes decreased and the expression of inflammatory factors increased, resulting in an increase in cellular apoptosis. Gp helped RGCs to become resistant to oxidation damage by directly reducing the amount of ROS in cells and exerting protective effects against H2O2-induced apoptosis. Treatment with Gp also reduced the generation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and increased nuclear respiratory factor 2 (Nrf-2) levels so as to increase the levels of heme oxygenase-1 (HO-1) and glutathione peroxidase 1/2 (Gpx1/2), which can enhance antioxidation in RGCs. In conclusion, our data indicate that neuroprotection by Gp involves its antioxidation and anti-inflammation effects. Gp prevents apoptosis through a mitochondrial apoptotic pathway. This finding might provide novel insights into understanding the mechanism of the neuroprotective effects of gypenosides in the treatment of optic neuritis.

Wolbachia supplement biotin and riboflavin to enhance reproduction in planthoppers.

Symbiont-mediated nutritional mutualisms can contribute to the host fitness of insects, especially for those that feed exclusively on nutritionally unbalanced diets. Here, we elucidate the importance of B group vitamins in the association of endosymbiotic bacteria Wolbachia with two plant-sap feeding insects, the small brown planthopper, Laodelphax striatellus (Fallén), and the brown planthopper, Nilaparvata lugens (Stål). Infected planthoppers of both species laid more eggs than uninfected planthoppers, while the experimental transfer of Wolbachia into uninfected lines of one planthopper species rescued this fecundity deficit. The genomic analysis showed that Wolbachia strains from the two planthopper species encoded complete biosynthesis operons for biotin and riboflavin, while a metabolic analysis revealed that Wolbachia-infected planthoppers of both species had higher titers of biotin and riboflavin. Furthermore, experimental supplementation of food with a mixture of biotin and riboflavin recovered the fecundity deficit of Wolbachia-uninfected planthoppers. In addition, comparative genomic analysis suggested that the riboflavin synthesis genes are conserved among Wolbachia supergroups. Biotin operons are rare in Wolbachia, and those described share a recent ancestor that may have been horizontally transferred from Cardinium bacteria. Our research demonstrates a type of mutualism that involves a facultative interaction between Wolbachia and plant-sap feeding insects involving vitamin Bs.

Traditional uses, phytochemistry, and pharmacology of Persicaria orientalis (L.) Spach - A review.

ETHNOPHARMACOLOGICAL RELEVANCE: Persicaria orientalis (L.) Spach (internationally accepted and only valid name; synonym: Polygonum orientale L.; family: Polygonaceae), which is named Hongcao in China, is a Chinese herbal medicine that has a wide range of pharmacological effects including treatment to rheumatoid arthritis, coronary heart disease, hernia, carbuncle sore, enhance immunity, antimicrobial, osteogenic and dilated bronchiectasis. AIM OF THIS REVIEW: This review aims to provide systematically organized information on traditional uses of Persicaria orientalis (L.) Spach (P. orientalis) and to critically analyze evidences in phytotherapeutic, botanical, and pharmacological literatures that support its therapeutic potential in treatment to human diseases. Isolation of additional compounds and detailed pharmacological investigations are key areas to investigate. MATERIALS AND METHODS: Relevant information on P. orientalis was collected through published scientific materials (including PubMed, ScienceDirect, Wiley, ACS, CNKI, Scifinder, Springer, Taylor & Francis, Web of Science, Google Scholar, and Baidu Scholar) and other literature sources (e.g., Chinese Pharmacopoeia, 2015 edition, Chinese herbal classic books and PhD and MSc thesis, etc.). RESULTS: Traditional uses were compiled in this review, including classic prescriptions and historical applications. Approximately 70 compounds, mainly including flavonoids, phenolics, lignans, limonoids and steroids, have been isolated and identified from P. orientalis. Among them, flavonoids were main components. Crude extracts and pure compounds isolated from P. orientalis exhibited various pharmacological activities, such as protection against ischemia and hypoxia-induced myocardial cells and hypoxia/reoxygenation cardiomyocyte, increase the blood flow in myocardium, expanding bronchus, anti-inflammatory and analgesic, and antithrombotic effects and so on. CONCLUSIONS: P. orientalis is a valuable source with therapeutic potential on a wide range of diseases especially cardiovascular-system disorders. Though most traditional uses of P. orientalis are supported by in vitro/vivo pharmacological studies, however, there is still a lack of researches on active pharmacodynamic ingredients as well as in-depth and in-vivo mechanistic studies. Therefore, isolation and identification of more active compounds (especially flavonoids), their structure-activity relationship and studies on pharmacodynamic mechanisms by more elaborative in-vivo studies on P. orientalis may be focused on in order to confirm efficacy of reported therapeutic effects of P. orientalis and help explore it's therapeutic potentials. Furthermore, research designs of pharmacological studies based on traditional uses of anti-rheumatoid arthritis through cell lines and animal models should also be considered as key research topics.

Chemical Constituents from Tabernaemontana bufalina Lour.

Investigation of the branches and leaves of Tabernaemontana bufalina Lour. led to afford an undescribed monoterpenoid indole alkaloid, namely (3R,7S,14R,19S,20R)-19-hydroxypseudovincadifformine (1), and nine known metabolites (2-10). Their structures were determined by analysis of their NMR and ESI-MS spectra, and modified Mosher's and calculated electronic circular dichroism (ECD) methods were used for establishing the absolute configuration of compound 1. Their cytotoxic activities were assayed using two cancer cell lines. As the results, cytotoxic activities on MDA-MB-231 and B16 cells showed IC50 values of 8.9 and 0.13 µm for 6, and of 20.3 and 11.7 µm for 9, respectively.

Anti-rheumatoid arthritis effects in adjuvant-induced arthritis in rats and molecular docking studies of Polygonum orientale L. extracts.

BACKGROUND AND PURPOSE: Polygonum orientale L. (family: Polygonaceae), named Hongcao in China, has effects of dispelling wind and dampness, promoting blood circulation, and relieving pain. Our group has already studied and confirmed that POEa and POEe (ethyl acetate and ethyl ether extract of P. orientale, respectively) had anti-inflammatory and analgesic effects in early research, which was mainly relevant to the existence of flavonoids. According to the clinical application of P. orientale in traditional Chinese medicine, it has long been used for rheumatic arthralgia and rheumatoid arthritis. Therefore, our group further explored whether flavonoids of P. orientale have anti-rheumatoid arthritis effect and how does they play this role. METHODS: Dried small pieces of the stems and leaves of P. orientale were decocted with water and partitioned successively to obtain POEa and POEe, respectively. The anti-rheumatoid arthritis effect of P. orientale was studied by using a Freund's complete adjuvant (FCA)-induced arthritis (AIA) in a rat model. The levels of PGE2, TNF-α, and IL-1ß in serum of AIA rats were detected by enzyme linked immunosorbent assay (ELISA) to explore its mechanisms. In addition, we computationally studied the relationships between the 15 chemical components of POEa and POEe, and the currently focused 9 target proteins of rheumatoid arthritis by molecular docking. RESULTS: Pharmacological experiments showed that POEa and POEe significantly ameliorate symptoms of rheumatoid arthritis via reducing paw swelling volume, arthritis score, and thymus and spleen indices, as well as increasing body weight in AIA rats. Simultaneously, the concentrations of PGE2, TNF-α, and IL-1ß were significantly decreased by POEa and POEe. Histopathology revealed noticeable reduction in bone and cartilage, synovial hyperplasia, inflammatory cell infiltration, cartilage surface erosion, and joint degeneration by POEa and POEe treatment. In addition, the molecular docking studies showed that docking scores of 14 chemical compositions (including 12 flavonoids and 2 phenolic acids) of POEa and POEe with anti-rheumatoid arthritis protein targets were better than the complexed ligands of the anti-rheumatoid arthritis protein targets. Among them, six flavonoids in POEa and POEe had more docking protein targets (n ≥ 3). Five anti-rheumatoid arthritis targets including high-temperature requirement A1 protease (HtrA1), janus kinase 1 (JAK1), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (i-NOS), and prostaglandin E2 (PGE2) had better docking score compared with the complexed ligands. Moreover, most of the chemical components in POEa and POEe showed strong interaction with HtrA1. CONCLUSIONS: The flavonoids of P. orientale have anti-rheumatoid arthritis effect. In addition, the molecular docking results indicate that quercetin, catechol, orientin, and other six flavonoids may be closely related to HtrA1, JAK1, COX-2, i-NOS, and PGE2 protein target receptors. It suggests that these chemical compositions form strong protein-ligand complexes with these protein targets, especially HtrA1 to exert anti-rheumatoid arthritis. Further experimental studies show that mechanisms of anti-rheumatoid arthritis effects may also be relevant to inhibit the levels of PGE2, TNF-α, and IL-1ß in serum. Therefore, our group can further explore the possible active ingredients and mechanisms of the anti-rheumatoid arthritis effects of flavonoids, and focus on the inhibition of the expression of inflammatory factors and the TGF-ß1/Smad signaling pathway associated with HtrA1 protein target receptors, which can provide a direction and powerful reference for the action mechanism and drug research of anti-rheumatoid arthritis of flavonoids in P. orientale.

Comparative metabolomics analysis for the compatibility and incompatibility of kansui and licorice with different ratios by UHPLC-QTOF/MS and multivariate data analysis.

Kansui, the root of Euphorbia kansui T.N. Liou ex T.P. Wang (Euphorbiaceae), is a well-known poisonous traditional Chinese medicine (TCM). However, many monographs of TCM indicated that it cannot be co-used with licorice, as kansui-licorice is a typical "eighteen incompatible" medicaments. Our previous studies have indicated that kansui was effective in treating malignant pleural effusion (MPE), and the efficacy could be weakened by the co-use of licorice, even causing serious toxicity at the given ratio. Nevertheless, the actual mechanisms of their dosage-toxicity-efficacy relationship need to be well clarified. The present study aimed to investigate the effect of individual and combined use of kansui and licorice on MPE rats, and explain the underlying mechanisms from a metabolomic perspective. Urine samples were analyzed by ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS). Partial least-squares discriminate analysis (PLS-DA) models were built to evaluate the interaction between kansui and licorice. Seven potential biomarkers contribute to the separation of model group and control group were tentatively identified. And selenoamino acid metabolism and nicotinate and nicotinamide metabolism with the impact-value 0.31 and 0.24, respectively, were filtered out as the most important metabolic pathways. Kansui and kansui-licorice at a ratio of 4:1 can treat MPE rats by adjusting abnormal metabolic pathways to the normal state, while it may have opposite result with kansui-licorice 1:4. The different influences to the two metabolic pathways may partially explain the dosage-toxicity-efficacy relationship of kansui-licorice with different ratios. The results could offer valuable insights into the compatibility property changes for the two herbs.

The Chemical and Biological Properties of Euphorbia kansui.

Kansui, the root of Euphorbia kansui T.N. Liou ex T.P. Wang, is a well-known traditional Chinese medicine. This paper reviews advances in investigations of the botany, the phytochemistry, the analytical method, the pharmacology and the toxicology of kansui. Nearly 100 compounds have been isolated from kansui and identified, and diterpenes and triterpenes are considered to be the characteristic and bio-active constituents of kansui. They possess multiple pharmacological activities, including diuretic, purgation, and antitumor effects. However, they also have a degree of toxicity, and can cause skin, oral, and gastrointestinal irritation. In this paper, the toxicity-efficacy relationship, attenuation and incompatibility of kansui are further discussed. Several future investigations of kansui are also proposed, all of which would improve the identification of kansui and other toxic herbs, as well as further their utilization.

The dosage-toxicity-efficacy relationship of kansui and licorice in malignant pleural effusion rats based on factor analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: The root of Euphorbia kansui T.P. Wang (Euphorbiaceae), a well-known traditional Chinese medicine (TCM) with certain toxicity, is known as Gan sui (Chinese: ) or kansui. It has been used to treat edema, ascites, asthma, and etc. Licorice is the root of Glycyrrhiza uralensis Fisch. or Glycyrrhiza inflate Bat. or Glycyrrhiza glabra L., Leguminosae. It is a widely used herbal medicine native to southern Europe and parts of Asia as an herbal medicine and natural sweetener. Kansui cannot be co-used with licorice, which is recorded in "eighteen incompatible" medicaments in many monographs of TCM. AIM OF THE STUDY: The present study was conducted to investigate the dosage-toxicity-efficacy relationship of the co-use of kansui and licorice and to explore its regularity of the toxicity and efficacy change. MATERIALS AND METHODS: Malignant pleural effusion rats were used and randomly divided into the normal control group, model group, positive control group (furosemide), kansui group, licorice group, and kansui-licorice groups with different ratios (kansui: licorice: 4:1, 2:1, 1:1, 0.5:1, 0.25:1, 0.1:1). Each group was adopted simultaneously to investigate the characteristic of toxicity and effect by measuring the pleural fluid and urine volumes, serum biochemical indexes, and serum TNF-α, IL-2 and IFN-γ levels. The factor analytic approach was used to analyze the dosage-toxicity-efficacy relationship between kansui and licorice. RESULTS: Two common factors were extracted from 8 indexes concerning toxicity and 5 indexes concerning efficacy. And the total factors related to toxicity (Ft) and efficacy (Fe) were calculated. The curved line of Ft indicated that the toxicity was increased along with the dose increase in licorice. The curved line of Fe indicated that the efficacy was decreased along with the dose increase in licorice. The intersection of these two lines was between the ratios of 2:1 and 1:1, and was deemed the flex point of the dosage-toxicity-efficacy. CONCLUSIONS: Kansui demonstrated a certain efficacy in treating malignant pleural effusion, and the efficacy could be weakened by the co-use of licorice, even causing serious toxicity at the given ratio. The ratio between 2:1 and 1:1 (kansui: licorice) was deemed the flex point of the dosage-toxicity-efficacy of kansui and licorice. The results will be helpful for their better utilization and development.

[Dosage-toxicity-efficacy relationship of Kansui Radix in malignant pleural effusion models based on Walker 256 analysis].

To investigate the dosage-toxicity-efficacy relationship of Kansui Radix, explore its regularity of the toxicity and efficacy change, and provide scientific basis for its clinical rational application, the malignant pleural effusion models were used to observe the effect of Kansui Radix with larger dose range (0.045-1.620 g•kg•d⁻¹ for Kansui Radix) on biochemical indexes and hydrothorax volume in experimental animals. Factor analysis method was also used to comprehensively assess the dosage-toxicity-efficacy relationship of Kansui Radix. The results showed that the rats in model group had larger hydrothorax volume, and ALT, AST, LDH, HBDH, IFN-γ, IL-2 and TNF-α levels were significantly increased (P<0.05), while TP and ALB levels were decreased (P<0.05) as compared with the blank group. After drug administration, various treatment groups decreased hydrothorax volume, IFN-γ, IL-2, TNF-α and increased TP and ALB levels as compared with model group, indicating certain therapeutic effect; and increased ALT, AST, LDH and HBDH levels, indicating certain liver and cardiac toxicity. In the factor analysis, two common factors were extracted from nine indexes, explaining 89.1% of the information. The analysis results suggested that there was no obvious toxicity in case of independent use of Kansui Radix within the dosage range set in pharmacopeia, while it would produce liver toxicity and cardiac toxicity upon 3 times of the dosage set in pharmacopeia, and the toxicity was increased with the increase of dose. At the same time, Kansui Radix can decrease the hydrothorax volume in malignant pleural effusion models and improve relevant physical indexes in a dose-dependent manner. Comprehensive analysis results of its toxic effect characteristics indicated that the upper-limit dose of Kansui Radix in pharmacopeia shall be regarded as the relatively optimal therapeutic dose.

Urine and plasma metabonomics coupled with UHPLC-QTOF/MS and multivariate data analysis on potential biomarkers in anemia and hematinic effects of herb pair Gui-Hong.

ETHNOPHARMACOLOGICAL RELEVANCE: The compatibility of Angelicae Sinensis Radix (Danggui) and Carthami Flos (Honghua), a famous herb pair Gui-Hong, can produce synergistic and complementary hematinic effects. Our previous studies have indicated that Gui-Hong has therapeutic potential treatment in hemolytic and aplastic anemia (HAA). The present study aimed to investigate the hematinic effects of Danggui, Honghua and Gui-Hong on HAA rats induced by acetyl phenylhydrazine (APH) and cyclophosphamide (CP) and to explore the underlying hematinic regulation mechanisms. MATERIALS AND METHODS: Rats were divided into 5 groups, and drugs were administered by oral gavage one time each day for continuous 7 days from the experiment began. Urine and plasma were analyzed by ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS). Partial least-squares discriminate analysis (PLS-DA) models were built to evaluate the therapeutic effects of Danggui, Honghua and Gui-Hong. Pearson correlation matrix analysis method was used to discover the correlations between potential biomarkers and biochemical indicators of HAA rats. RESULTS: Seven potential biomarkers contribute to the separation of model group and control group were tentatively identified. The levels of l-kynurenine, phenylalanine, nicotinic acid and sphingosine increased significantly (P<0.05) in HAA rats, while the levels of l-isoleucine, l-tyrosine and serotonin decreased significantly (P<0.05) in comparison with control rats. Those endogenous metabolites were chiefly involved in phenylalanine, tyrosine and tryptophan biosynthesis, valine, leucine and isoleucine biosynthesis, tryptophan metabolism and tyrosine metabolism. The metabolic deviations could be regulated closer to normal level after Danggui, Honghua and Gui-Hong intervention. In term of hematinic effects, Gui-Hong was the most effective as shown by the relative distance in PLS-DA score plots and relative intensity of potential biomarkers. The result reflected the synergic action between Danggui and Honghua. The above results were found to be reasonable in explaining the hematinic effects mechanism of Gui-Hong. CONCLUSIONS: The results of routine blood, urinary metabolic pattern and plasma metabolic pattern show the Danggui, Honghua and Gui-Hong groups are moving toward the control group and the HAA was being prevented and alleviated. The effect of Gui-Hong group is more remarkable than Danggui and Honghua groups. Some potential biomarkers like l-kynurenine, phenylalanine, l-isoleucine, l-tyrosine, serotonin, nicotinic acid and sphingosine have been found and identified. The work shows that the metabonomics method is a promising tool in the efficacy and mechanism research of traditional Chinese medicines.

Comparative metabolomics analysis on invigorating blood circulation for herb pair Gui-Hong by ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry and pattern recognition approach.

The compatibility of Angelicae Sinensis Radix (Danggui, DG) and Flos Carthami (Honghua, HH), a famous herb pair Gui-Hong (GH), can produce synergistic and promoting blood effects. Although some physiological and pathological function parameters of the acute blood stasis have been investigated, little information about the changes of small metabolites in biofluids has been reported. In present study, global metabolic profiling with ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS) combined with pattern recognition method was performed to discover the underlying blood-activating regulation mechanisms of DG, HH and GH on the acute blood stasis rats induced by subcutaneous injection of adrenaline hydrochloride and ice water bath. The total 14 metabolites (10 in urine and 4 in plasma), up regulated or down regulated (P<0.05 or 0.01), were identified and contributed to the acute blood stasis progress. These promising identified biomarkers underpin the metabolic pathway including phenylalanine metabolism, sphingolipid metabolism, arachidonic acid metabolism and arginine and proline metabolism are disturbed in the acute blood stasis rats, which identified by using pathway analysis with MetPA. The altered metabolites and hemorheological indexes could be regulated closer to normal level after DG, HH and GH intervention. In term of activate blood circulation function, GH was the most effective as shown by the relative distance in PLS-DA score plots and relative intensity of metabolomics trategy, reflecting the synergic action between Danggui and Honghua. The results demonstrated that biofluids metabolomics was a powerful tool in clinical diagnosis and treatment of syndrome of blood stasis for providing information on changes in metabolites pathways.

Anti-allodynic effects of obtusifolin and gluco-obtusifolin against inflammatory and neuropathic pain.

Inflammatory pain and neuropathic pain are major health issues that represent considerable social and economic burden worldwide. In this study we investigated the potential of obtusifolin and gluco-obtusifolin, two anthraquinones found in the seeds of the widely used traditional Chinese medical botanical Cassia obtusifolia, to reduce neuropathic and inflammatory pain. The potential analgesic effects of obtusifolin and gluco-obtusifolin were evaluated by mice formalin test and complete Freund's adjuvant (CFA)-induced nociceptive behaviors in rats. Chronic constriction injury (CCI), L5 spinal nerve ligation (L5 SNL), diabetes, and chemotherapeutics inducing allodynia were used to test whether repeated treatment with obtusifolin and gluco-obtusifolin ameliorated neuropathic pain. Finally, we explored whether obtusifolin and gluco-obtusifolin altered the degree of neuroinflammation in rat spinal cord after CFA administration and CCI induction. Obtusifolin and gluco-obtusifolin (0.25, 0.5, 1, and 2 mg/kg) reduced licking/biting time in dose-dependent manner in phase 2 of formalin-induced behavior in mice. Furthermore, repeated administration of obtusifolin and gluco-obtusifolin (0.25, 0.5, 1, and 2 mg/kg) reversed mechanical allodynia induced by CFA, CCI, L5 SNL, diabetes, and oxaliplatin in a dose-dependent manner in rats. Levels of activated nuclear factor-kappa B (NF-κB) and proinflammatory cytokines (interleukin (IL)-1ß, IL-6, tumor necrosis factor α (TNF-α)) in lumbar spinal cord were elevated in rats following CFA treatment and CCI induction, and obtusifolin and gluco-obtusifolin significantly inhibited these effects. Our results demonstrate that obtusifolin and gluco-obtusifolin produce significant antinociceptive action in rodent behavioral models of inflammatory/neuropathic pain, and that this activity is associated with modulation of neuroinflammation in spinal cord.

Medicinal plants of the genus Gelsemium (Gelsemiaceae, Gentianales)--a review of their phytochemistry, pharmacology, toxicology and traditional use.

ETHNOPHARMACOLOGICAL RELEVANCE: In the genus Gelsemium, Gelsemium elegans (Gardn. & Champ.) Benth. has been recognized as a toxic plant that is widely distributed in Southeast Asia and has been used as traditional Chinese medicine for the treatment of rheumatoid pain, neuropathic pain, spasticity, skin ulcers and cancers for many years. Gelsemium sempervirens (L.) J.St.-Hil. has been used since the nineteenth century in homeopathy for treating anxiety, neuralgia, migraine and spasmodic disorders, such as asthma and whooping cough in North America. This review aims to provide comprehensive information on the botany, traditional uses, phytochemistry, pharmacological research and toxicology of medicinal plants in the genus Gelsemium. The overall objective is to explore the evidence supporting its ethnopharmacological effectiveness. MATERIALS AND METHODS: A literature survey was performed by searching the scientific databases Pubmed, Google Scholar, SciFinder, Scopus, Web of Science and the Chinese CNKI, in addition to traditional Chinese medicine and homeopathic texts for information on Gelsemium. RESULTS: Plants of the genus Gelsemium have been used in traditional medicine for the treatment of migraines, neuralgia, sciatica, cancer and various types of sores. Studies into the phytochemical composition of this genus have shown that all of the species are rich sources of monoterpene indole alkaloids and that they have attracted the attention of many researchers due to their markedly diverse and complex architecture. To date, a total of 121 alkaloids have been isolated and identified from the genus. The crude extracts, as well as the monomeric compounds, from the genus possess anti-tumor, analgesic, anxiolytic, anti-inflammatory and immunomodulating pharmacological activities. CONCLUSION: It is evident from the available literature that Gelsemium species possess potential for use as a beneficial therapeutic remedy. However, the analysis of previous pharmacological research suggests that a clear assignment of active molecules and mechanisms of action is remain lacking. Due to their high toxicity, the studies available on toxicity and safety are inadequate for providing information on clinical utilization.