SLC25A3 negatively regulates NLRP3 inflammasome activation by restricting the function of NLRP3.

The NACHT, leucine-rich repeat, and pyrin domains-containing protein 3 (collectively known as NLRP3) inflammasome activation plays a critical role in innate immune and pathogenic microorganism infections. However, excessive activation of NLRP3 inflammasome will lead to cellular inflammation and tissue damage, and naturally it must be precisely controlled in the host. Here, we discovered that solute carrier family 25 member 3 (SLC25A3), a mitochondrial phosphate carrier protein, plays an important role in negatively regulating NLRP3 inflammasome activation. We found that SLC25A3 could interact with NLRP3, overexpression of SLC25A3 and knockdown of SLC25A3 could regulate NLRP3 inflammasome activation, and the interaction of NLRP3 and SLC25A3 is significantly boosted in the mitochondria when the NLRP3 inflammasome is activated. Our detailed investigation demonstrated that the interaction between NLRP3 and SLC25A3 disrupted the interaction of NLRP3-NEK7, promoted ubiquitination of NLRP3, and negatively regulated NLRP3 inflammasome activation. Thus, these findings uncovered a new regulatory mechanism of NLRP3 inflammasome activation, which provides a new perspective for the therapy of NLRP3 inflammasome-associated inflammatory diseases.

KIAA1549 promotes the development and chemoresistance of colorectal cancer by upregulating ERCC2.

Colorectal cancer (CRC) is the third most commonly diagnosed cancer worldwide. Chemotherapy is the mainstay of treatment for patients with CRC in II-IV stages. Resistance to chemotherapy occurs commonly, which results in treatment failure. Therefore, the identification of novel functional biomarkers is essential for recognizing high-risk patients, predicting recurrence, and developing new therapeutic strategies. Herein, we assessed the roles of KIAA1549 in promoting tumor development and chemoresistance in colorectal cancer. As a result, we found that KIAA1549 expression is up-regulation in CRC. Public databases revealed a progressive up-regulation of KIAA1549 expression from adenomas to carcinomas. Functional characterization uncovered that KIAA1549 promotes tumor malignant phenotypes and boosts the chemoresistance of CRC cells in an ERCC2-dependent manner. Inhibition of KIAA1549 and ERCC2 effectively enhanced the sensitivity to chemotherapeutic drugs oxaliplatin and 5-fluorouracil. Our findings suggest that endogenous KIAA1549 might function as a tumor development-promoting role and trigger chemoresistance in colorectal cancer partly by upregulating DNA repair protein ERCC2. Hence, KIAA1549 could be an effective therapeutic target for CRC and inhibition of KIAA1549 combined with chemotherapy might be a potential therapeutic strategy in the future.

DENV NS1 and MMP-9 cooperate to induce vascular leakage by altering endothelial cell adhesion and tight junction.

Dengue virus (DENV) is a mosquito-borne pathogen that causes a spectrum of diseases including life-threatening dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Vascular leakage is a common clinical crisis in DHF/DSS patients and highly associated with increased endothelial permeability. The presence of vascular leakage causes hypotension, circulatory failure, and disseminated intravascular coagulation as the disease progresses of DHF/DSS patients, which can lead to the death of patients. However, the mechanisms by which DENV infection caused the vascular leakage are not fully understood. This study reveals a distinct mechanism by which DENV induces endothelial permeability and vascular leakage in human endothelial cells and mice tissues. We initially show that DENV2 promotes the matrix metalloproteinase-9 (MMP-9) expression and secretion in DHF patients' sera, peripheral blood mononuclear cells (PBMCs), and macrophages. This study further reveals that DENV non-structural protein 1 (NS1) induces MMP-9 expression through activating the nuclear factor κB (NF-κB) signaling pathway. Additionally, NS1 facilitates the MMP-9 enzymatic activity, which alters the adhesion and tight junction and vascular leakage in human endothelial cells and mouse tissues. Moreover, NS1 recruits MMP-9 to interact with ß-catenin and Zona occludens protein-1/2 (ZO-1 and ZO-2) and to degrade the important adhesion and tight junction proteins, thereby inducing endothelial hyperpermeability and vascular leakage in human endothelial cells and mouse tissues. Thus, we reveal that DENV NS1 and MMP-9 cooperatively induce vascular leakage by impairing endothelial cell adhesion and tight junction, and suggest that MMP-9 may serve as a potential target for the treatment of hypovolemia in DSS/DHF patients.

Integration of glucose and cardiolipin anabolism confers radiation resistance of HCC.

BACKGROUND AND AIMS: Poor response to ionizing radiation (IR) due to resistance remains a clinical challenge. Altered metabolism represents a defining characteristic of nearly all types of cancers. However, how radioresistance is linked to metabolic reprogramming remains elusive in hepatocellular carcinoma (HCC). APPROACH AND RESULTS: Baseline radiation responsiveness of different HCC cells were identified and cells with acquired radio-resistance were generated. By performing proteomics, metabolomics, metabolic flux, and other functional studies, we depicted a metabolic phenotype that mediates radiation resistance in HCC, whereby increased glucose flux leads to glucose addiction in radioresistant HCC cells and a corresponding increase in glycerophospholipids biosynthesis to enhance the levels of cardiolipin. Accumulation of cardiolipin dampens the effectiveness of IR by inhibiting cytochrome c release to initiate apoptosis. Mechanistically, mammalian target of rapamycin complex 1 (mTORC1) signaling-mediated translational control of hypoxia inducible factor-1α (HIF-1α) and sterol regulatory element-binding protein-1 (SREBP1) remodels such metabolic cascade. Targeting mTORC1 or glucose to cardiolipin synthesis, in combination with IR, strongly diminishes tumor burden. Finally, activation of glucose metabolism predicts poor response to radiotherapy in cancer patients. CONCLUSIONS: We demonstrate a link between radiation resistance and metabolic integration and suggest that metabolically dismantling the radioresistant features of tumors may provide potential combination approaches for radiotherapy in HCC.

Vortex array generation based on quasi-Talbot effects.

A lens-less method for generating vortex arrays with tunable parameters is proposed based on quasi-Talbot effects. By illuminating a two-dimensional periodic sinusoidal grating with a vortex beam carrying a fourth-order cross-phase, the continuous vortex array structure can be generated in the Fresnel diffraction region. Due to the shaping effect of the fourth-order cross-phase on the vortex beam, by changing the constant parameter of the fourth-order cross-phase, it is possible to shape the generation of optical vortex arrays at different positions. This will somewhat broaden the flexibility of the lens-free optical vortex array in terms of generation position. In addition, the generation of polygonal optical vortex arrays is achieved by higher-order cross-phases of different orders. This technique has potential applications in various fields such as optical tweezers, multi-particle screening, microscopic manipulation, etc.

TXN inhibitor impedes radioresistance of colorectal cancer cells with decreased ALDH1L2 expression via TXN/NF-κB signaling pathway.

BACKGROUND: Colorectal cancer (CRC) is prevalent worldwide and is often challenged by treatment failure and recurrence due to resistance to radiotherapy. Here, we aimed to identify the elusive underlying molecular mechanisms of radioresistance in CRC. METHODS: Weighted gene co-expression network analysis was used to identify potential radiation-related genes. Colony formation and comet assays and multi-target single-hit survival and xenograft animal models were used to validate the results obtained from the bioinformatic analysis. Immunohistochemistry was performed to examine the clinical characteristics of ALDH1L2. Co-immunoprecipitation, immunofluorescence and flow cytometry were used to understand the molecular mechanisms underlying radioresistance. RESULTS: Bioinformatic analysis, in vitro, and in vivo experiments revealed that ALDH1L2 is a radiation-related gene, and a decrease in its expression induces radioresistance in CRC cells by inhibiting ROS-mediated apoptosis. Patients with low ALDH1L2 expression exhibit resistance to radiotherapy. Mechanistically, ALDH1L2 interacts with thioredoxin (TXN) and regulates the downstream NF-κB signaling pathway. PX-12, the TXN inhibitor, overcomes radioresistance due to decreased ALDH1L2. CONCLUSIONS: Our results provide valuable insights into the potential role of ALDH1L2 in CRC radiotherapy. We propose that the simultaneous application of TXN inhibitors and radiotherapy would significantly ameliorate the clinical outcomes of patients with CRC having low ALDH1L2.

STING promotes NLRP3 localization in ER and facilitates NLRP3 deubiquitination to activate the inflammasome upon HSV-1 infection.

One of the fundamental reactions of the innate immune responses to pathogen infection is the release of pro-inflammatory cytokines, including IL-1ß, processed by the NLRP3 inflammasome. The stimulator of interferon genes (STING) has the essential roles in innate immune response against pathogen infections. Here we reveal a distinct mechanism by which STING regulates the NLRP3 inflammasome activation, IL-1ß secretion, and inflammatory responses in human cell lines, mice primary cells, and mice. Interestingly, upon HSV-1 infection and cytosolic DNA stimulation, STING binds to NLRP3 and promotes the inflammasome activation through two approaches. First, STING recruits NLRP3 and facilitates NLRP3 localization in the endoplasmic reticulum, thereby facilitating the inflammasome formation. Second, STING interacts with NLRP3 and attenuates K48- and K63-linked polyubiquitination of NLRP3, thereby promoting the inflammasome activation. Collectively, we demonstrate that the cGAS-STING-NLRP3 signaling is essential for host defense against HSV-1 infection.

Research on ACEI of Low-Molecular-Weight Peptides from Hirudo nipponia Whitman.

The renin-angiotensin system (RAS) is the primary pathway for regulating blood pressure in the body, and angiotensin-converting enzymes (ACEs) play a crucial role in it. Hirudo nipponia is an invertebrate that contains a variety of active peptides; however, there are no studies on the ACE inhibitory activity of hirudo. In the present study, our aim was to identify the active peptides in hirudo based on active peptide database analysis, unexpectedly filling the gap in hirudo ACE inhibitory activity research. Prep-HPLC was used to separate the part below 3 kD from hirudo. The peptide composition of the isolates was obtained based on Orbitrap LC-MS. The activity of each group of peptides was predicted by the database and the activity was determined by bioassay. Peptides with validation activity were screened through the database. In total, 337 peptides and 18 peptides matching the NCBI leech protein database were identified. All four fractions showed ACE inhibitory activity, and the IC50 was 0.8266, 0.2708, 0.4432, and 0.1764 mg/mL, respectively. Six screened peptides showed good affinity for ACE. This work reveals for the first time that low-molecular-weight peptides from H. nipponia have ACE inhibitory activity, which can provide a new explanation for leech treatment of hypertension.

Paxillin mediates ATP-induced activation of P2X7 receptor and NLRP3 inflammasome.

BACKGROUND: Extracellular adenosine triphosphate (ATP), a key danger-associated molecular pattern (DAMP) molecule, is released to the extracellular medium during inflammation by injured parenchymal cells, dying leukocytes, and activated platelets. ATP directly activates the plasma membrane channel P2X7 receptor (P2X7R), leading to an intracellular influx of K+, a key trigger inducing NLRP3 inflammasome activation. However, the mechanism underlying P2X7R-mediated activation of NLRP3 inflammasome is poorly understood, and additional molecular mediators have not been identified. Here, we demonstrate that Paxillin is the molecule connecting the P2X7 receptor and NLRP3 inflammasome through protein interactions. RESULTS: We show a distinct mechanism by which Paxillin promotes ATP-induced activation of the P2X7 receptor and NLRP3 inflammasome. Extracellular ATP induces Paxillin phosphorylation and then facilitates Paxillin-NLRP3 interaction. Interestingly, Paxillin enhances NLRP3 deubiquitination and activates NLRP3 inflammasome upon ATP treatment and K+ efflux. Moreover, we demonstrated that USP13 is a key enzyme for Paxillin-mediated NLRP3 deubiquitination upon ATP treatment. Notably, extracellular ATP promotes Paxillin and NLRP3 migration from the cytosol to the plasma membrane and facilitates P2X7R-Paxillin interaction and PaxillinNLRP3 association, resulting in the formation of the P2X7R-Paxillin-NLRP3 complex. Functionally, Paxillin is essential for ATP-induced NLRP3 inflammasome activation in mouse BMDMs and BMDCs as well as in human PBMCs and THP-1-differentiated macrophages. CONCLUSIONS: We have identified paxillin as a mediator of NLRP3 inflammasome activation. Paxillin plays key roles in ATP-induced activation of the P2X7 receptor and NLRP3 inflammasome by facilitating the formation of the P2X7R-Paxillin-NLRP3 complex.

Histone methyltransferase SETDB1 promotes colorectal cancer proliferation through the STAT1-CCND1/CDK6 axis.

Upregulation of histone methyltransferase SET domain bifurcated 1 (SETDB1) is associated with poor prognosis in cancer patients. However, the mechanism of oncogenicity of SETDB1 in cancer is hitherto unknown. Here, we show that SETDB1 is upregulated in human colorectal cancer (CRC) where its level correlates with poor clinical outcome. Ectopic SETDB1 promotes CRC cell proliferation, whereas SETDB1 attenuation inhibits this process. Flow cytometry reveals that SETDB1 promotes proliferation by driving the CRC cell cycle from G0/G1 phase to S phase. Mechanistically, SETDB1 binds directly to the STAT1 promoter region resulting in increased STAT1 expression. Functional characterization reveals that STAT1-CCND1/CDK6 axis is a downstream effector of SETDB1-mediated CRC cell proliferation. Furthermore, SETDB1 upregulation is sufficient to accelerate in vivo proliferation in xenograft animal model. Taken together, our results provide insight into the upregulation of SETDB1 within CRC and can lead to novel treatment strategies targeting this cell proliferation-promoting gene.

Runoff simulation of two typical urban green land types with the Stormwater Management Model (SWMM): sensitivity analysis and calibration of runoff parameters.

The characteristics of surface runoff and the infiltration properties of urban green land are important to determine the effects of runoff reduction by low-impact development (LID) facilities. In this paper, two typical types of urban green land (lawn and shrub) in Shanghai were selected to study the runoff characteristics under eight rainfall events. The sensitivity of the runoff parameters was analyzed, and then, the optimal parameters were determined using the Stormwater Management Model (SWMM). The results showed that the interception and infiltration capacities of shrub were greater than those of lawn. The rainfall intensity and rainfall pattern were the major factors that influenced the interception and infiltration of rainwater. The threshold value that generates runoff varied across the eight rainfall events ranged from 1.6 to 28.5 mm for lawn and 4.5 to 32.0 mm for shrub. The maximum reduction ratios of runoff and peak flow for shrub were 52 and 57% higher than them for lawn, respectively. The parameters for shrub were more sensitive to runoff and peak flow compared with those for lawn. Under light rainfalls with a short duration, the maximum infiltration rate and depression storage were more sensitive than those under heavy rainfalls with a long duration. Antecedent dry weather period was not found to be a sensitive parameter except for the shrub under light rainfalls. The relative errors of runoff and dynamic mean runoff (60 min) for lawn and shrub were within ± 9.5%. The errors of peak flow ranged between - 21 and 16.6%. The dynamic runoff characteristics and the parameters for lawn and shrub determined in this study can provide references for simulating urban runoff and planning LID areas.

Histone methyltransferase SETDB1 promotes cells proliferation and migration by interacting withTiam1 in hepatocellular carcinoma.

BACKGROUND: SETDB1 is a histone H3K9 methyltransferase, which plays a significant role in the occurrence and progression of tumors. Previous studies have confirmed that T-lymphom invasion and metastasis gene (Tiam1) is a protein associated with the metastasis of hepatocellular carcinoma (HCC); however, we have not yet been successful in elucidating the specific mechanism of HCC. METHODS: Yeast two-hybrid test was conducted to screen proteins that interacted with Tiam1 gene. Glutathione-S-transferase (GST) pull-down and crosslinking-immunoprecipitation (CLIP) assays were performed to determine whether SETDB1 can interact with Tiam1 gene. A series of related experiments were performed to explore role of SETDB1 on cell proliferation, migration, and invasion in HCC. Recovery experiment was performed to investigate the effect of Tiam1 knockdown on cell proliferation and migration, which was caused by SETDB1 overexpression in HCC cells. The expression of SETDB1 was frequently upregulated in HCC tissues and positively correlated with Tiam1. RESULTS: GST pull-down and CLIP assays were performed to elucidate the interaction between SETDB1 and Tiam1. Cell proliferation, migration, and epithelial mesenchymal transformation (EMT) in HCC cells was promoted with the overexpression of SETDB1. Following the knockdown of Tiam1 gene, the effect of SETDB1 on cell proliferation and migration was reversed in HCC cells. The expression of SETDB1 was frequently up-regulated in HCC tissues, and it was positively correlated with Tiam1 gene. CONCLUSIONS: Ours is the first study to prove that SETDB1 promotes the proliferation and migration of cells by forming SETDB1-Tiam1 compounds. We found that SETDB1-Tiam1 compounds were involved in a novel pathway, which regulated epigenetic modification of gene expression in HCC samples.

Different erosion characteristics of sediment deposits in combined and storm sewers.

To investigate the different erosion patterns of sediments in combined and storm sewers, sediments from three separate sewer systems and two combined sewer systems in urban Shanghai were collected for the flushing experiments. These experiments were conducted with different consolidation periods and shear velocities. As the consolidation period increases, dissolved oxygen exhibits a positive effect on the microbial transformations of organic substrates. Potential structural changes and separations of the surface and bottom layers of sediments are observed. The results also reveal that the organic matter, particle size and moisture have different effects on the erosion resistance of sediments. Furthermore, illicit connections behaved as an important factor affecting the viscosity and static friction force of particles, which directly alter the erosion resistance of sewer sediments.

[Study on formation and changes of Dao-di herbs production origin of Gastrodia elata].

Gastrodia elata has been used in China for more than 2 000 years and it is a kind of valuable traditional Chinese medicine. The originrecords of G. elata were Mount Tai of Shandong and and Mount Song of Henan, which began in Wupu Bencao of Wei Jin Dynasties, and Tai'an and its surrounding areas had been the Do-di herbs production areas. But from the beginning of the Republic of China, G. elata origin has undergone major changes, Do-di herbs production areas moved westward to the southwest.In this paper,through literature research and field visits, we studied the formation and changes of Do-di herbs production areas of G. elata. The cultivation history and current main producing area of G. elata was also introduced. On this basis, we profoundly summarized the reasons of Do-di herbs production areas formation and changes from the nature, society, transportation, humanities and germplasm resources.Combining the ancient herbal medicine and the characteristics of modern producing areas, the planting strength of G. elata could be strengthened in the hope of providing reference for the quality evaluation and cultivation of G. elata.

Low Temperature Soda-Oxygen Pulping of Bagasse.

Wood shortages, environmental pollution and high energy consumption remain major obstacles hindering the development of today's pulp and paper industry. Energy-saving and environmental friendly pulping processes are still needed, especially for non-woody materials. In this study, soda-oxygen pulping of bagasse was investigated and a successful soda-oxygen pulping process for bagasse at 100 °C was established. The pulping parameters of choice were under active alkali charge of 23%, maximum cooking temperature 100 °C, time hold at maximum temperature 180 min, initial pressure of oxygen 0.6 MPa, MgSO4 charge 0.5%, and de-pithed bagasse consistency 12%. Properties of the resultant pulp were screened yield 60.9%, Kappa number 14, viscosity 766 dm³/kg, and brightness 63.7% ISO. Similar pulps were also obtained at 110 °C or 105 °C with a cooking time of 90 min. Compared with pulps obtained at higher temperatures (115-125 °C), this pulp had higher screened yield, brightness, and acceptable viscosity, while the delignification degree was moderate. These results indicated that soda-oxygen pulping at 100 °C, the lowest cooking temperature reported so far for soda-oxygen pulping, is a suitable process for making chemical pulp from bagasse. Pulping at lower temperature and using oxygen make it an environmental friendly and energy-saving pulping process.

[Induction and analysis for NIR features of frequently-used mineral traditional Chinese medicines].

In order to provide theoretical basis for the rapid identification of mineral traditional Chinese medicines(TCM) with near infrared (NIR)diffuse reflectance spectroscopy, Characteristic NIR spectra of 51 kinds of mineral TCMs were generalized and compared on the basis of the previous research, and the characteristic spectral bands were determined and analyzed by referring to mineralogical and geological literatures. It turned out that the NIR features of mineral TCMs were mainly at 8 000-4 000 cm ⁻¹ wavebands, which can be assigned as the absorption of water, -OH and[CO3 ²â»] and so on. Absorption peaks of water has regularity as follows, the structure water and -OH had a combined peak which was strong and keen-edged around 7 000 cm ⁻¹, the crystal water had two strong peak around 7 000 cm ⁻¹ and 5 100 cm ⁻¹, and water only has a broad peak around 5 100 cm ⁻¹. Due to the differences in the crystal form and the contents of water in mineral TCMs, NIR features of water in mineral TCMs which could be used for identification were different. Mineral TCMs containing sulfate are rich in crystal water, mineral TCMs containing silicate generally had structure water, and mineral TCMs containing carbonate merely had a little of water, so it was reasonable for the use of NIR spectroscopy to classify mineral TCMs with anionic type. In addition, because of the differences in cationic type, impurities, crystal form and crystallinity, mineral TCMs have exclusive NIR features at 4 600-4 000 cm ⁻¹, which can be assigned as Al-OH, Mg-OH, Fe-OH, Si-OH,[CO3 ²â»] and so on. Calcined mineral TCMs are often associated with water and main composition changes, also changes of the NIR features, which could be used for the monitoring of the processing, and to provide references for the quality control of mineral TCMs. The adaptability and limitation of NIR analysis for mineral TCMs were also discussed:the majority of mineral TCMs had noteworthy NIR features which could be used for the NIR analysis. And the NIR features of a few mineral TCMs were inapparent, such as Fluoritum, Realgar and Cinnabar, for which the Raman spectroscopy can be adopted alternatively.

[Analysis on 24 Samples of Fluoritum by XRD Pattern].

Objective: To provide the reference for effective identification of Fluoritum,by using X-ray diffraction technique to analyze Fluoritum samples which had different morphological features. Methods: According to the China Pharmacopeia( 2010 edition),the24 samples of commercial Fluoritum were identified and their contents of Ca F2 were determined. XRD technique was applied to analyze phase compositions and content from Fluoritum samples, to ensure quality, and to summarize the correlation between traits and quality. Results: Sample 1 ~ 7 and 13 were Fluoritum, samples 8 ~ 12 were inferior products which were doped, and sample 14 ~ 24 were counterfeit products. Fluorite was the main phase of Fluoritum, and often accompanying a small amount of quartz. Phase compositions of counterfeit Fluoritum whose impurity content were high were relatively complicated, and the contents of Ca F2 were far below the standards value of the China Pharmacopeia( 2010 edition). Fluoritum were easy to be shattered into tiny sand which were green or purple, hyaline and lustered, with color becoming shallow. Conclusion: By picking to remove impurity, inferior products can be used for medicine. Because impurity content are high and the impurities are difficult to be separated, counterfeit products can not be used for medicine. Characteristics of powder can be used for supplement the identification for Fluoritum. And using XRD technique can accurately identify Fluoritum samples which have different morphological features.

[Identification of Mineral Medicine of Chloriti Lapis,Micae Lapis Aureus and Vermiculitum by X-ray Diffraction].

Objective: To analyze the mineral medicine of Chloriti Lapis, Micae Lapis Aureus and Vermiculitum by X-ray diffraction, then to guide the identification and quality assessment of them. Methods: XRD Fourier patterns were collected from powder samples to analyze phase compositions, and to determine the original mineral resources of Chloriti Lapis, Micae Lapis Aureus and Vermiculitum by comparing with their characteristic traits. First derivative + vector normalization and 21 point smoothing were used to pretreat the selected spectrum band from 0. 68 ~ 1. 77 nm. Then the data were analyzed by fuzzy cluster. Results: It was found that the original mineral resource of seven powder samples of Chloriti Lapis was biotite schist belonging to metamorphic mineral. The original mineral resource of three powder samples of high-quality Micae Lapis Aureus was vermiculite biotite schist belonging to metamorphic mineral. The original mineral resources of three powder samples of Vermiculitum were phlogopite and vermiculite phlogopite. Conclusion: The method of X-ray diffraction analysis is accurate and rapid, which can be used for the identification and quality evaluation of Chloriti Lapis, Micae Lapis Aureus and Vermiculitum.

[Taxonomy and Distribution of Chinese Medicinal Centipedes].

Objective: To study the taxonomy and distribution of Chinese medicinal centipedes. Methods: The species of Chinese medicinal centipedes were investigated in the light of their morphology. According to the feature of life, the distribution of centipedes were explored. Results: There were 12 centipede species in China, and seven of them were used for medical, the species could be effectively distinguished by the identification key. It was suggested that their characteristics were related to the climatic factors such as temperature, humidity, altitude and air pressure. Conclusion: The distribution of medicinal centipede is characteristics of "three river system distribution belts" and "three geographical distribution areas". The results provide the basis for the development and application of medicinal centipedes in China.

Separation and purification of four flavonol diglucosides from the flower of Meconopsis integrifolia by high-speed counter-current chromatography.

Flavonoids are the main components of Meconopsis integrifolia (Maxim.) Franch, which is a traditional Tibetan medicine. However, traditional chromatography separation requires a large quantity of raw M. integrifolia and is very time consuming. Herein, we applied high-speed counter-current chromatography in the separation and purification of flavonoids from the ethanol extracts of M. integrifolia flower. Ethyl acetate/n-butanol/water (2:3:5, v/v/v) was selected as the optimum solvent system to purify the four components, namely quercetin-3-O-ß-d-glucopyrannosy-(1→6)-ß-d-glucopyranoside (compound 1, 60 mg), quercetin 3-O-[2'''-O-acetyl-ß-d-glucopyranosyl-(1→6)-ß-d-glucopyranoside (compound 2, 40 mg), quercetin 3-O-[3'''-O-acetyl-ß-d-glucopyranosyl-(1→6)-ß-d-glucopyranoside (compound 3, 11 mg), and quercetin 3-O-[6'''-O-acetyl-ß-d-glucopyranosyl-(1→6)-ß-d-glucopyranoside (compound 4, 16 mg). Among the four compounds, 3 and 4 were new acetylated flavonol diglucosides. After the high-speed counter-current chromatography separation, the purities of the four flavonol diglucosides were 98, 95, 90, and 92%, respectively. The structures of these compounds were identified by mass spectrometry and NMR spectroscopy.