Adsorption effect and mechanism of Cd(II) by different phosphorus-enriched biochars.

The resource utilization of agricultural and forestry waste, especially the high-value transformation of low-grade phosphate rock and derivatives, is an important way to achieve sustainable development. This study focuses on the impregnation and co-pyrolysis of rice straw (RS) with fused calcium magnesium phosphate (FMP), FMP modified with citric acid (CA-FMP), and calcium dihydrogen phosphate (MCP) to produce three phosphorous-enriched biochars (PBC). The Cd(II) removal efficiency of biochars before and after phosphorus modification was investigated, along with the adsorption mechanism and contribution of biochars modified with different phosphorus sources to Cd(II) adsorption. The result indicated that CA-FMP and MCP could be more uniformly loaded onto biochar, effectively increasing the specific surface area (SSA) and total pore volume. The adsorption of Cd(II) onto PBC followed a mono-layer chemisorption process accompanied by intraparticle diffusion. The adsorption of Cd(II) by PBC involved ion exchange, mineral precipitation, complexation with oxygen-containing functional groups (OFGs), cation-π interaction, electrostatic interaction, and physical adsorption. Ion exchange was identified as the primary adsorption mechanism for Cd(II) by BC and FBC (51.53% and 53.15% respectively), while mineral precipitation played a major role in the adsorption of Cd(II) by CBC and MBC (51.10% and 47.98% respectively). Moreover, CBC and MBC significantly enhanced the adsorption capacity of Cd(II), with maximum adsorption amounts of 128.1 and 111.5 mg g-1 respectively.

Methyl donor diet attenuates intimal hyperplasia after vascular injury in rats.

Environmental factors, particularly dietary habits, play an important role in cardiovascular disease susceptibility and progression through epigenetic modification. Previous studies have shown that hyperplastic vascular intima after endarterectomy is characterized by genome-wide hypomethylation. The purpose of this study was to investigate whether methyl donor diet affects intimal hyperplasia and the possible mechanisms involved. Intimal hyperplasia was induced in SD rats by carotid artery balloon injury. From 8 d before surgery to 28 d after surgery, the animals were fed a normal diet (ND) or a methyl donor diet (MD) supplemented with folic acid, vitamin B12, choline, betaine, and zinc. Carotid artery intimal hyperplasia was observed by histology, the effect of MD on carotid protein expression was analyzed by proteomics, functional clustering, signaling pathway, and upstream-downstream relationship of differentially expressed proteins were analyzed by bioinformatics. Results showed that MD attenuated balloon injury-induced intimal hyperplasia in rat carotid arteries. Proteomic analysis showed that there were many differentially expressed proteins in the common carotid arteries of rats fed with two different diets. The differentially expressed proteins are mainly related to the composition and function of the extracellular matrix (EMC), and changes in the EMC can lead to vascular remodeling by affecting fibrosis and stiffness of the blood vessel wall. Changes in the levels of vasculotropic proteins such as S100A9, ILF3, Serpinh1, Fbln5, LOX, HSPG2, and Fmod may be the reason why MD attenuates intimal hyperplasia. Supplementation with methyl donor nutrients may be a beneficial measure to prevent pathological vascular remodeling after injury.

Selenium regulates T cell differentiation in experimental autoimmune thyroiditis in mice.

Selenium (Se) is an essential trace element that plays an important role in thyroid physiology. Se supplementation can reduce levels of autoimmune thyroid antibodies, which may be beneficial in Hashimoto's thyroiditis (HT). However, the long-term benefits of Se supplementation for HT patients are controversial and there is no clear clinical evidence to support it, so further basic and clinical research is needed. The effect of Se on immune cells, especially T cells, in autoimmune thyroiditis (AIT) has not been elucidated. Here, we replicated a mouse model of experimental autoimmune thyroiditis (EAT) on a high-iodine diet and treated it with Se supplementation. At week 8 of the experiment, Se supplementation reduced the destruction of thyroid follicles and the infiltration rate of lymphocytes in EAT mice, and reversed the disturbance of peripheral blood thyroxine and thyroid autoantibody levels. Further examination revealed that Se had broad effects on T-cell subsets. Its effects include reducing the production of pro-inflammatory cytokines by Th1 cells, inhibiting the differentiation and production of cytokines by Th2 and Th17 cells, and upregulating the differentiation and production of cytokines by Treg cells. These changes help alleviate thyroid follicle damage during EAT. In conclusion, selenium supplementation has the potential to improve the prognosis of AIT by altering the subset differentiation and/or function of CD4+ T cells.

Thyrotropin inhibits osteogenic differentiation of human periodontal ligament stem cells.

BACKGROUND AND OBJECTIVE: Periodontal ligament stem cells (PDLSCs) are derived from the periodontal ligament and have the characteristics of pluripotent differentiation, including osteogenesis, and are one of the important seed cells in oral tissue engineering. Thyrotropin (TSH) has been shown to regulate bone metabolism independently of thyroid hormone, including the fate of osteoblasts and osteoclasts, but whether it affects osteogenic differentiation of PDLSCs is unknown. MATERIALS AND METHODS: PDLSCs were isolated and cultured from human periodontal ligament and grown in osteogenic medium (containing sodium ß-glycerophosphate, ascorbic acid, and dexamethasone). Recombinant human TSH was added to the culture medium. Osteogenic differentiation of PDLSCs was assessed after 14 days by staining with alkaline phosphatase and alizarin red and by detection of osteogenic differentiation genes. Differentially expressed genes (DEGs) in PDLSCs under TSH were detected by high-throughput sequencing. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyzed the biological functions and signaling pathways involved in DEGs. RESULTS: We found that osteogenic differentiation of PDLSCs was significantly inhibited in the presence of TSH: including decreased calcium nodule formation, decreased alkaline phosphatase levels, and decreased collagen synthesis. Using high-throughput sequencing, we found changes in the expression of some osteogenesis-related genes, which may be the reason that TSH inhibits osteogenic differentiation of PDLSCs. CONCLUSION: Unless TSH is ≥10 mU/L, patients with subclinical hypothyroidism usually do not undergo thyroxine supplementation therapy. However, in this work, we found that elevated TSH inhibited the osteogenic differentiation of PDLSCs. Therefore, correction of TSH levels in patients with subclinical hypothyroidism may be beneficial to improve orthodontic, implant, and periodontitis outcomes in these patients.

Predicting the Stability of Organic Matter Originating from Different Waste Treatment Procedures.

Recycling organic wastes into farmland faces a double challenge: increasing the carbon storage of soil while mitigating CO2 emission from soil. Predicting the stability of organic matter (OM) in wastes and treatment products can be helpful in dealing with this contradiction. This work proposed a modeling approach integrating an OM characterization protocol into partial least squares (PLS) regression. A total of 31 organic wastes, and their products issued from anaerobic digestion, composting, and digestion-composting treatment were characterized using sequential extraction and three-dimension (3D) fluorescence spectroscopy. The apportionment of carbon in different fractions and fluorescence spectra revealed that the OM became less accessible and biodegradable after treatments, especially the composting. This was proven by the decrease in CO2 emission from soil incubation. The PLS model successfully predicted the stability of solid digestate, compost, and compost of solid digestate in the soil by using only the characterized variables of non-treated wastes. The results suggested that it would be possible to predict the stability of OM from organic wastes after different treatment procedures. It is helpful to choose the most suitable and economic treatment procedure to stabilize labile organic carbon in wastes and hence minimize CO2 emission after the application of treatment products to the soil.

Transcriptome Analysis Provides Insights into Potentilla bifurca Adaptation to High Altitude.

Potentilla bifurca is widely distributed in Eurasia, including the Tibetan Plateau. It is a valuable medicinal plant in the Tibetan traditional medicine system, especially for the treatment of diabetes. This study investigated the functional gene profile of Potentilla bifurca at different altitudes by RNA-sequencing technology, including de novo assembly of 222,619 unigenes from 405 million clean reads, 57.64% of which were annotated in Nr, GO, KEGG, Pfam, and Swiss-Prot databases. The most significantly differentially expressed top 50 genes in the high-altitude samples were derived from plants that responded to abiotic stress, such as peroxidase, superoxide dismutase protein, and the ubiquitin-conjugating enzyme. Pathway analysis revealed that a large number of DEGs encode key enzymes involved in secondary metabolites, including phenylpropane and flavonoids. In addition, a total of 298 potential genomic SSRs were identified in this study, which provides information on the development of functional molecular markers for genetic diversity assessment. In conclusion, this study provides the first comprehensive assessment of the Potentilla bifurca transcriptome. This provides new insights into coping mechanisms for non-model organisms surviving in harsh environments at high altitudes, as well as molecular evidence for the selection of superior medicinal plants.

The complete chloroplast genome of a medical herb, Rheum lhasaense (Polygonaceae), from Qinghai-Tibet Plateau in China.

Rheum lhasaense (Polygonaceae) is one of the genuine medicinal herbs in Qinghai-Tibet Plateau, China. Here we report the first chloroplast (cp) genome of R. lhasaense using Illumina NovaSeq 6000 platform. The length of its complete cp genome is 161,820 bp, containing four sub-regions. A large single copy region (LSC) of 87,086 bp and a small single copy region (SSC) of 12,814 bp are separated by a pair of inverted repeat regions (IRs) of 30,960 bp. The complete cp genome of R. lhasaense contains 130 genes, including 85 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The overall GC content of the cp genome is 37.4%. The phylogenetic analysis, based on 28 cp genomes, suggested that R. lhasaense is closely related to R. acuminatum and R. pumilum.

The combined therapy of a traditional Chinese medicine formula and Western medicine for a critically ill case infected with COVID-19.

OBJECTIVE: Presentation of a case illustrating the benefits of traditional Chinese medicine (TCM) for treatment of Coronavirus disease 2019 (COVID-19) in critically ill patients. CLINICAL FEATURES AND OUTCOME: A 58-year-old woman presented with cough, fever, dizziness, chest tightness, polypnea and poor appetite. She was admitted to Guizhou Provincial People's hospital, and diagnosed with critically ill type of COVID-19 in February 2020. According to the patient's symptoms and signs, the TCM syndrome differentiation was qi deficiency, dampness-stasis and toxin accumulation. Then she received the combined therapy of a modified Chinese herbal formula and Western medicine. During a twelve-day period of treatment, her respiratory distress and appetite quickly improved. Abnormal laboratory indicators were resumed in time and lung lesions in CT scan largely absorbed. No side effects associated with this Chinese herbal formula were found. Before discharge, two consecutive nasopharyngeal swabs were shown to be negative for severe acute respiratory coronavirus 2 (SARS-CoV-2). CONCLUSIONS: Our case report suggests that collaborative treatments with traditional Chinese medicine prove beneficial in the management of COVID-19 in critically ill patients. In order to give optimal care for this COVID-19 crisis for the whole world, Chinese medicine practitioners and Western medical doctors should work together in frontline.

Effects of bioporous carriers on the performance and microbial community structure in side-stream anaerobic membrane bioreactors.

The aim of this study was to investigate the effects of a volcanic rock porous carrier (VRPC) on sludge reduction, pollutant removal, and microbial community structure in an anaerobic side-stream reactor (ASSR). Three lab-scale membrane bioreactors (MBRs), including an anoxic-oxic MBR, which served as the control (C-MBR), an ASSR-coupled MBR (A-MBR), and an A-MBR filled with VRPC (FA-MBR) were stably and simultaneously operated for 120 days. The effect of the three reactors on the removal of chemical oxygen demand (COD) was almost negligible (all greater than 95%), but the average removal efficiency of ammonium nitrogen, total nitrogen, and total phosphorus was significantly improved by the insertion of an ASSR, especially when the ASSR was filled with VRPC. Finally, A-MBR and FA-MBR achieved 16.2% and 26.4% sludge reduction rates, with observed sludge yields of 0.124 and 0.109 g mixed liquid suspended solids/g COD, respectively. Illumina MiSeq sequencing revealed that microbial diversity and richness were highest in the VRPC, indicating that a large number of microorganisms formed on the carrier surface in the form of a biofilm. Abundant denitrifying bacteria (Azospira, Comamonadaceae_unclassified, and Flavobacterium) were immobilized on the carrier biofilm, which contributed to increased nitrogen removal. The addition of a VRPC to the ASSR successfully immobilized abundant hydrolytic, fermentative, and slow-growing microorganisms, which all contributed to reductions in sludge yield.

A new methylene bisflavan-3-ol from the branches and leaves of Potentilla fruticosa.

Dasiphora fruticosa L. (Rosaceae), also known as Potentilla fruticosa L. (syn.), is a hardy deciduous shrub widely distributed in the north temperate regions of the world. Three methylene bisflavan-3-ols (1-3), together with a procyanidin dimer, (-)-afzelechin-(4α→8)-(-)-afzelechin (4) were isolated for the first time from the branches and leaves of the titled plant, in addition to 11 known compounds (5-15). Their structures were elucidated by means of extensive spectroscopic analysis and by comparison with data reported in the literatures. Methylene 6,8-bis(7-O-glucosyl) catechin (1) was determined to be a new dimeric flavan-3-ol glycoside through a methylene linkage between C-8 and C-8 of two units. At a concentration of 128 µg/mL, the known compounds 9 - 13 exhibited antibacterial activities on Escherichia coli, Staphylococcus aureus subsp. aureus, Salmonella enterica subsp. enterica, and Pseudomonas aeruginosa. Compound 12 also showed certain glucose uptake stimulating activity.

The impact of giant jellyfish Nemopilema nomurai blooms on plankton communities in a temperate marginal sea.

This study focused on the bloom-developing process of the giant jellyfish, Nemopilema nomurai, on phytoplankton and microzooplankton communities. Two repeated field observations on the jellyfish bloom were conducted in June 2012 and 2014 in the southern Yellow Sea where blooms of N. nomurai were frequently observed. We demonstrated that the bloom was made up of two stages, namely the developing stage and the mature stage. Total chlorophyll a increased and the concentrations of inorganic nutrients decreased during the developing stage, while both concentrations maintained stable and at low levels during the mature stage. Our analysis revealed that phosphate excreted by growing N. nomurai promoted the growth of phytoplankton at the developing stage. At the mature stage, size compositions of microzooplankton were altered and tended to be smaller via a top-down process, while phytoplankton compositions, affected mainly through a bottom-up process, shifted to be less diatoms and cryptophytes but more dinoflagellates.

Warming and eutrophication combine to restructure diatoms and dinoflagellates.

Temperature change and eutrophication are known to affect phytoplankton communities, but relatively little is known about the effects of interactions between simultaneous changes of temperature and nutrient loading in coastal ecosystems. Here we show that such interaction is key in driving diatom-dinoflagellate dynamics in the East China Sea. Diatoms and dinoflagellates responded differently to temperature, nutrient concentrations and ratios, and their interactions. Diatoms preferred lower temperature and higher nutrient concentrations, while dinoflagellates were less sensitive to temperature and nutrient concentrations, but tended to prevail at low phosphorus and high N:P ratio conditions. These different traits of diatoms and dinoflagellates resulted in the fact that both the effect of warming resulting in nutrients decline as a consequence of increasing stratification and the effect of increasing terrestrial nutrient input as a result of eutrophication might promote dinoflagellates over diatoms. We predict that conservative forecasts of environmental change by the year 2100 are likely to result in the decrease of diatoms in 60% and the increase of dinoflagellates in 70% of the surface water of the East China Sea, and project that mean diatoms should decrease by 19% while mean dinoflagellates should increase by 60% in the surface water of the coastal East China Sea. This analysis is based on a series of statistical niche models of the consequences of multiple environmental changes on diatom and dinoflagellate biomass in the East China Sea based on 2815 samples randomly collected from 23 cruises spanning 14 years (2002-2015). Our findings reveal that dinoflagellate blooms will be more frequent and intense, which will affect coastal ecosystem functioning.

A urinary metabolomics (GC-MS) strategy to evaluate the antidepressant-like effect of chlorogenic acid in adrenocorticotropic hormone-treated rats.

Major depressive disorder (MDD) is a chronic recurring illness that seriously affects human health. Chlorogenic acid (CGA), an important polyphenol extracted from Eucommia ulmoides Oliver bark, has been reported to have anti-depression, neuroprotection, memory improvement and other pharmacological effects. However, little is known about the underlying mechanisms of CGA on the treatment of depression. Here, we investigated the antidepressant-like effects of CGA on an adrenocorticotropic hormone (ACTH)-treated rat model. Thirty-two male Wistar rats were randomly divided into four groups: normal diet group (N), ACTH-treated model group (M), memantine positive control group (M + Mem) and CGA intervened group (M + CGA). Sucrose preference tests (SPTs) and open-field tests (OFTs) were performed to evaluate depressive-like behaviors. Memantine (30 mg kg-1) and CGA (500 mg kg-1) administration dramatically increased hedonic behaviors of the rats in SPT. The scores of crossing and rearing were significantly increased in the M + Mem group and M + CGA group. These results of the behaviour tests might be suggestive of antidepressant-like effects. Moreover, memantine and CGA reversed the levels of serum 5-hydroxytryptamine (5-HT), ACTH, corticotropin-releasing hormone (CRH), and dopamine (DA) that were altered in ACTH-treated rats. Based on a GC-MS metabolomic approach, significant differences in the metabolic profile were observed in ACTH-treated rats compared with the control group, as well as the M + CGA group and M + Mem group compared with the ACTH-treated group. A total of 19 metabolites were identified for the discrimination of normal rats and ACTH-treated rats, and 12 out of 19 differential metabolites were reversed with CGA intervention. Combined with pattern recognition and bioinformatics, nine perturbed metabolic pathways, including energy metabolism, neurotransmitter metabolism, and amino acid metabolism, were identified based on these metabolites. These integrative studies might give a holistic insight into the pathophysiological mechanism of the ACTH-treated depressive rat model, and also showed that CGA has antidepressant-like activities in ACTH-treated rats, providing an important drug candidate for the prevention and treatment of tricyclic anti-depressant treatment-resistant depression.

Semiconductor diode laser device adjuvanting intradermal vaccine.

A brief exposure of skin to a low-power, non-tissue damaging laser light has been demonstrated to augment immune responses to intradermal vaccination. Both preclinical and clinical studies show that this approach is simple, effective, safe and well tolerated compared to standard chemical or biological adjuvants. Until now, these laser exposures have been performed using a diode-pumped solid-state laser (DPSSL) devices, which are expensive and require labor-intensive maintenance and special training. Development of an inexpensive, easy-to-use and small device would form an important step in translating this technology toward clinical application. Here we report that we have established a handheld, near-infrared (NIR) laser device using semiconductor diodes emitting either 1061, 1258, or 1301nm light that costs less than $4000, and that this device replicates the adjuvant effect of a DPSSL system in a mouse model of influenza vaccination. Our results also indicate that a broader range of NIR laser wavelengths possess the ability to enhance vaccine immune responses, allowing engineering options for the device design. This small, low-cost device establishes the feasibility of using a laser adjuvant approach for mass-vaccination programs in a clinical setting, opens the door for broader testing of this technology with a variety of vaccines and forms the foundation for development of devices ready for use in the clinic.

Generation of human MHC (HLA-A11/DR1) transgenic mice for vaccine evaluation.

The rapid occurrence of emerging infectious diseases demonstrates an urgent need for a new preclinical experimental model that reliably replicates human immune responses. Here, a new homozygous humanized human leukocyte antigen (HLA)-A11/DR1 transgenic mouse (HLA-A11(+/+)/DR01(+/+)/H-2-ß2m(-/-)/IAß(-/-)) was generated by crossing HLA-A11 transgenic (Tg) mice with HLA-A2(+/+)/DR01(+/+)/H-2-ß2m(-/-)/IAß(-/-) mice. The HLA-A11-restricted immune response of this mouse model was then examined. HLA-A11 Tg mice expressing a chimeric major histocompatibility complex (MHC) molecule comprising the α1, α2, and ß2m domains of human HLA-A11 and the α3 transmembrane and cytoplasmic domains of murine H-2D(b) were generated. The correct integration of HLA-A11 and HLA-DR1 into the genome of the HLA-A11/DR1 Tg mice (which lacked the expression of endogenous H-2-I/II molecules) was then confirmed. Immunizing mice with a recombinant HBV vaccine or a recombinant HIV-1 protein resulted in the generation of IFN-γ-producing cytotoxic T lymphocyte (CTL) and antigen-specific antibodies. The HLA-A11-restricted CTL response was directed at HLA immunodominant epitopes. These mice represent a versatile animal model for studying the immunogenicity of HLA CTL epitopes in the absence of a murine MHC response. The established animal model will also be useful for evaluating and optimizing T cell-based vaccines and for studying differences in antigen processing between mice and humans.

Improving composting as a post-treatment of anaerobic digestate.

This work investigated the influences of practical parameters upon composting of digestate. The yardsticks for evaluation were digestate stabilization, nitrogenous emissions mitigation and self-heating potential. The results suggest choosing an "active" bulking agent like dry wood chips (WC) which served as free-water and nitrogen sink through composting. At an optimal volumetric WC:digestate mixing ratio of 4:1, nearly 90% of the initial NH4(+)/NH3 were fixed, which reduced significantly nitrogenous emissions. This mixing ratio also improved the stabilization and self-heating potential. Using small particle size WC increased narrowly O2 consumption and reduced NH3 emission. Storing used WC prior to recycling reduced 40% N2O emission compared to directly recycled WC. Recycling compost helped to decrease NH3 emission, but quadrupled N2O emission. The optimal aeration rate (15Lh(-1)kg OM0) which was lower compared to composting of organic waste, was enough to ensure the O2 supply and ameliorate the self-heating potential through composting of digestate.

Ethanol extract of Dendrobium chrysotoxum Lindl ameliorates diabetic retinopathy and its mechanism.

Diabetic retinopathy (DR) is the most common and serious complication of diabetes mellitus (DM). The present study investigates the amelioration of ethanol extract of Dendrobium chrysotoxum Lindl (DC) on streptozotocin (STZ)-induced DR and its engaged mechanism. Retinal immunofluorescence staining with cluster of differentiation 31 (CD31) demonstrated that DC (30-300 mg/kg) decreased the increased retinal vessels in STZ-induced diabetic rats. Retinal histopathological observation also showed that retinal vessels were decreased in DC-treated diabetic rats. DC decreased the increased retinal mRNA expression of vascular endothelial growth factor (VEGF) and VEGF receptor 2 (VEGFR2) in diabetic rats, and DC also decreased the elevated serum VEGF level. Immunohistochemical staining further evidenced that DC decreased VEGF and VEGFR2 expression in retinas. Retinal mRNA expression of matrix metalloproteinase (MMP) 2/9 was decreased in DC (300 mg/kg)-treated diabetic rats. Serum levels of MMP 2/9, basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF) A/B, insulin-like growth factor 1 (IGF-1), interleukin 1ß (IL-1ß), and IL-6 were all decreased in DC-treated diabetic rats. In addition, DC decreased the increased phosphorylation of p65 and the increased expression of intercellular adhesion molecule-1 (ICAM-1). In conclusion, DC can alleviate retinal angiogenesis during the process of DR via inhibiting the expression of VEGF/VEGFR2, and some other pro-angiogenic factors such as MMP 2/9, PDGF A/B, bFGF, IGF-1. In addition, DC can also ameliorate retinal inflammation via inhibiting NFκB signaling pathway.

[Effects of Hippophae rhamnoides L. subsp. chinensis Rousi polysaccharide on alpha-glucosidase enzyme activity and level of blood glucose].

The enzyme-inhibitor model and the sugar tolerance mouse model were used to evaluate the relationship between the inhibition rate of enzyme activity and concentration of Hippophae rhamnoides L. subsp. chinensis Rousi polysaccharide (HRP). The inhibitory patterns of enzyme and dose-dependent effects of HRP's effect on blood glucose using acarbose tablets as control were also examined. The mechanism underlying hypoglycemic effects of HRP was discussed. The results showed: in the enzyme-inhibitor model, the inhibitory activity of different concentrations of HRP (9.80, 19.60, 39.20, 78.40, 156.80 and 312.50 mg x L(-1)) on alpha-glucosaminidase (AG) inhibitory activity were 6.62%, 18.02%, 33.26%, 48.23%, 62.11%, 76.31%, 90.12%, IC50 was 31.59 mg x L(-1). The inhibitory rate of 25.00 x 10(3) mg x L(-1) acarbose tablets was only 64.87%, and IC50 was 10.75 x 10(3) mg x L(-1). In the sugar tolerance mouse model, different doses of HRP (240, 480, 960 mg x kg(-1)) tended to decrease levels of blood glucose compared with control group (acarbose tablets 375 mg x kg(-1)) at 15, 30, 60 and 120 min. It's further confirmed that HRP is a kind of competitive inhibitor of AG activity. Its inhibition rate increases with the increase of concentration in normal mice, and it subsequently improves the sugar tolerance showing the effect of reducing blood sugar.

Anti-inflammatory compounds of "Qin-Jiao", the roots of Gentiana dahurica (Gentianaceae).

ETHNOPHARMACOLOGICAL RELEVANCE: "Qin-Jiao" is a well-known traditional Chinese medicinal (TCM) herb having been used generally for fighting rheumatoid arthritis (RA) since ancient times. The root of Gentiana dahurica Fisch (Gentianaceae) is one of the four officially validated "Qin-Jiao" as listed in the Chinese Pharmacopoeia. In addition, it is a common Tibetan medicinal herb used for the treatment of tonsillitis, urticaria, and RA, while the flowers have been used as a Mongolian herb for curing cough sore throat and eliminating the phlegm due to its anti-inflammatory effect. AIM OF THE STUDY: The aim of the study was to characterize the anti-inflammatory compounds in "Qin-Jiao", on the basis of detailed investigation on not only the phytochemical study of Gentiana dahurica, but also the bioactive evaluation on compounds obtained presently and previously from different "Qin-Jiao" origins and Gentiana species. MATERIALS AND METHODS: The ethanol extract of air-dried roots of Gentiana dahurica was suspended into H2O and extracted with EtOAc and n-BuOH, successively. Repeated column chromatography (CC) and semi-preparative HPLC were carried out on each of the fractions. The isolated compounds were determined by detailed spectroscopic analysis and acidic hydrolysis. Anti-inflammatory activities of 18 isolates, together with 12 typical compounds obtained previously by our group from the other "Qin-Jiao" origins (Gentiana crassicaulis, Gentiana straminea) and Gentiana rigescens, were tested by inhibitory effects on LPS-induced NO production in macrophage RAW264.7 cells and TPA-induced cyclooxygenases-2 and -1 (COXs-2/1) production on zebrafish model. RESULTS: A new lignan glycoside (1) was identified, together with 20 known compounds, including 10 iridoid glycosides (2-11), three steroids (12-14), four lignans (15-18), one phenylpropanoid (19) and two triterpenes (20-21). Anti-inflammatory bioassay showed that only compound 21 displayed potential inhibitory effect on NO production (IC50=16.85 µM), while 20 tested compounds had inhibitory activities on COXs-2/1. Among them, the triterpenoid 21 was the most active compound with an inhibitory value of 78% at a concentration of 30 µM. All the tested compounds showed no cytotoxicity on five human cancer cell lines (40µM) and zebrafish (30 µM), except for 21 displaying weak cytotoxicity on human myeloid leukemia HL-60 (IC50=16.43 µM). CONCLUSION: Most of compounds particularly iridoid glycosides from "Qin-Jiao" display potential inhibitory effect on COXs-2/1. The results support the historical importance of the well-known TCM herb, "Qin-Jiao", having been commonly used for fighting RA. As major components, the bioactive iridoid glycosides should play important role in the anti-inflammatory effect of "Qin-Jiao". Although further research will be required to evaluate the selective activities of the COXs-2/1 inhibitors, this work validates the medicinal use of "Qin-Jiao" and provides information for different "Qin-Jiao" origins having different treating effects on RA.