Sinomenine ameliorates collagen-induced arthritis in mice by targeting GBP5 and regulating the P2X7 receptor to suppress NLRP3-related signaling pathways.

Sinomenine (SIN) is an isoquinoline alkaloid isolated from Sinomenii Caulis, a traditional Chinese medicine used to treat rheumatoid arthritis (RA). Clinical trials have shown that SIN has comparable efficacy to methotrexate in treating patients with RA but with fewer adverse effects. In this study, we explored the anti-inflammatory effects and therapeutic targets of SIN in LPS-induced RAW264.7 cells and in collagen-induced arthritis (CIA) mice. LPS-induced RAW264.7 cells were pretreated with SIN (160, 320, 640 µM); and CIA mice were administered SIN (25, 50 and 100 mg·kg-1·d-1, i.p.) for 30 days. We first conducted a solvent-induced protein precipitation (SIP) assay in LPS-stimulated RAW264.7 cells and found positive evidence for the direct binding of SIN to guanylate-binding protein 5 (GBP5), which was supported by molecular simulation docking, proteomics, and binding affinity assays (KD = 3.486 µM). More importantly, SIN treatment markedly decreased the expression levels of proteins involved in the GBP5/P2X7R-NLRP3 pathways in both LPS-induced RAW264.7 cells and the paw tissue of CIA mice. Moreover, the levels of IL-1ß, IL-18, IL-6, and TNF-α in both the supernatant of inflammatory cells and the serum of CIA mice were significantly reduced. This study illustrates a novel anti-inflammatory mechanism of SIN; SIN suppresses the activity of NLRP3-related pathways by competitively binding GBP5 and downregulating P2X7R protein expression, which ultimately contributes to the reduction of IL-1ß and IL-18 production. The binding specificity of SIN to GBP5 and its inhibitory effect on GBP5 activity suggest that SIN has great potential as a specific GBP5 antagonist.

Study of the Mechanism of Action of Guanxin Shutong Capsules in the Treatment of Coronary Heart Disease Based on Metabolomics.

Background: Guan-Xin-Shu-Tong capsule (GXSTC) is a traditional Chinese medicine (TCM) that has been used to treat coronary heart disease (CHD) for many years in China. However, the holistic mechanism of GXSTC against CHD is still unclear. Therefore, the purpose of this paper was to systematically explore the mechanism of action GXSTC in the treatment of CHD rats using a metabolomics strategy. Methods: A CHD model was induced by ligation of the left anterior descending coronary artery (LAD). In each group, echocardiography was performed; the contents of creatine kinase (CK), lactate dehydrogenase (LDH) and aspartate transaminase (AST) in serum were determined; and the myocardial infarct size was measured. The metabolites in plasma were analyzed by UHPLC-MS/MS-based untargeted metabolomics. Then, multivariate statistical analysis was performed to screen potential biomarkers associated with the GXSTC treatment in the LAD-induced rat CHD model. Finally, the MetaboAnalyst 4.0 platform was used for metabolic pathway enrichment analysis. Results: GXSTC was able to regulate the contents of CK, LDH and AST; restore impaired cardiac function; and significantly reduce the myocardial infarction area in model rats. Twenty-two biomarkers and nine metabolic pathways of GXSTC in the treatment of CHD were identified through UHPLC-MS/MS-based untargeted metabolomics analysis. Conclusion: GXSTC regulates metabolic disorders of endogenous components in LAD-induced CHD rats. The anti-CHD mechanism of GXSTC is mainly related to the regulation of amino acid, lipid and hormonal metabolism. This study provides an overall view of the mechanism underlying the action of GXSTC against CHD.

Corrigendum to "Neuroprotective effect of Sanqi Tongshuan Tablets on sequelae post-stroke in rats" [Chinese Herbal Medicines 11 (2019) 45-51].

[This corrects the article DOI: 10.1016/j.chmed.2018.12.002.].

Targeted next-generation sequencing identified novel mutations in triple-negative myeloproliferative neoplasms.

Mutations in JAK2, MPL and CALR genes have been identified in the majority of myeloproliferative neoplasm (MPN) patients, and patients negative for these three mutations are the so-called triple-negative (TN) MPN. In this study, we examined the mutational profiles of 16 triple-negative MPN patients including 7 essential thrombocythemia (ET), 1 primary myelofibrosis and 8 polycythemia vera (PV). Targeted next-generation sequencing was performed using the ACTOnco Comprehensive Cancer Panel (Ion AmpliSeq Comprehensive Cancer Panel, Life Technologies) to target all coding exons of 409 cancer-related genes. Overall, 30 nonsynonymous somatic mutations were detected in 12 (75%) patients with a range of 1-5 mutations per sample. Notably, one ET patient was found to have JAK2V617F and KITP551L mutations at very low allele frequency. One MPLP70L and 1 MPLM602T mutations were identified each in 1 ET and 1 PV, respectively. Other recurrent mutations were also identified including KMT2C, KMT2D, IRS2, SYNE1, PDE4DIP, SETD2, ATM, TNFAIP3 and CCND2. In addition, germline mutations were also found in some cancer-related genes. Copy number changes were rare in this cohort of TN MPNs. In conclusion, both somatic and germline mutations can be detected in TN MPN patients.

Renoprotective Effects of Shout Camphor Medicinal Mushroom (Taiwanofungus camphorates, Basidiomycetes) Mycelia on Several Media in Mice with Chronic Kidney Disease.

Taiwanofungus camphoratus has been widely used in Taiwan as a folk medicine to prevent and treat liver diseases, diarrhea, abdominal pain, itchy skin, and hypertension. Recent studies have shown that T. camphoratus mycelia extracts exert anti-inflammatory and antioxidant effects on some types of renal disease, but the effect of T. camphoratus mycelia on chronic kidney disease (CKD) remains unclear. In this study we used the Bioresource Collection and Research Center (BCRC) medium and modified media (e.g., BCRC+A, HKS1, and HKS1+A media) to incubate T. camphorates mycelia and detect the feasible benefits of renal protection in mice with CKD. Five groups of mice with a partial nephrectomy (each mouse weighed approximately 30 g) received a daily administration of different media-treated T. camphoratus mycelia water solutions (3 mg dried mycelia dissolved in 0.3 mL water) by oral gavage for 30 days, while a control group received distilled water. The results show that progressive increased blood urea nitrogen and serum creatinine were significantly inhibited in the HKS1+A group on days 10 and 30. Plasma total protein was effectively increased in the HKS1 and HKS1+A groups. The BCRC and BCRC+A groups exhibited no obvious improvement in renal function. The results suggest that the HKS1+A medium provides the optimal effect in preventing the deterioration of kidney function and might have a renoprotective effect on CKD.

[Chemical Constituents of Euphorbia lunulata].

OBJECTIVE: To investigate the chemical constituents in the ethanol extract from the whole plant of Euphorbia lunulata. METHODS: The whole plant of Euphorbia lunulata was extracted by 95% ethanol, then partitioned by system solvents with different polarity. The ethyl acetate and n-butyl alcohol extracts were separated on silica gel, Sephadex LH-20,and MCI columns. The isolated compounds were determined by detailed analysis of their spectral data. RESULTS: Twelve compounds were isolated and identified from the ethyl acetate and n-butyl alcohol extracts of Euphorbia lunulata and the structures were identified as 7ß-methoxy-stigmast-5-ene-3ß-ol (1), 7ß-methoxy-stigmast-5-ene-3ß,22ß-diol(2), asperglaucide(3), moscatin (4), p-hydroxybenzoic acid (5),3-methoxy-4-hydroxy benzoic acid(6), erigeside C(7),5,7,4'-trihydroxy flavanone(8), kaempferol(9), quercetin(10), corosolic acid(11) and acacetin (12). CONCLUSION: All compounds except for 9 and 10 are reported from this plant for the first time.

A strategy that iteratively retains informative variables for selecting optimal variable subset in multivariate calibration.

Nowadays, with a high dimensionality of dataset, it faces a great challenge in the creation of effective methods which can select an optimal variables subset. In this study, a strategy that considers the possible interaction effect among variables through random combinations was proposed, called iteratively retaining informative variables (IRIV). Moreover, the variables are classified into four categories as strongly informative, weakly informative, uninformative and interfering variables. On this basis, IRIV retains both the strongly and weakly informative variables in every iterative round until no uninformative and interfering variables exist. Three datasets were employed to investigate the performance of IRIV coupled with partial least squares (PLS). The results show that IRIV is a good alternative for variable selection strategy when compared with three outstanding and frequently used variable selection methods such as genetic algorithm-PLS, Monte Carlo uninformative variable elimination by PLS (MC-UVE-PLS) and competitive adaptive reweighted sampling (CARS). The MATLAB source code of IRIV can be freely downloaded for academy research at the website: http://code.google.com/p/multivariate-calibration/downloads/list.

Synthesis and characterization of oil-chitosan composite spheres.

Oil-chitosan composite spheres were synthesized by encapsulation of sunflower seed oil in chitosan droplets, dropping into NaOH solution and in situ solidification. Hydrophilic materials (i.e., iron oxide nanoparticles) and lipophilic materials (i.e., rhodamine B or epirubicin) could be encapsulated simultaneously in the spheres in a one step process. The diameters of the prepared spheres were 2.48 ± 0.11 mm (pure chitosan spheres), 2.31 ± 0.08 mm (oil-chitosan composites), 1.49 ± 0.15 mm (iron-oxide embedded oil-chitosan composites), and 1.69 ± 0.1 mm (epirubicin and iron oxide encapsulated oil-chitosan composites), respectively. Due to their superparamagnetic properties, the iron-oxide embedded oil-chitosan composites could be guided by a magnet. A lipophilic drug (epirubicin) could be loaded in the spheres with encapsulation rate measured to be 72.25%. The lipophilic fluorescent dye rhodamine B was also loadable in the spheres with red fluorescence being observed under a fluorescence microscope. We have developed a novel approach to an in situ process for fabricating oil-chitosan composite spheres with dual encapsulation properties, which are potential multifunctional drug carriers.

Toxicity of areca nut ingredients: activation of CHK1/CHK2, induction of cell cycle arrest, and regulation of MMP-9 and TIMPs production in SAS epithelial cells.

BACKGROUND: There are 600 million betel quid chewers around the world. betel quid chewing is a major risk factor of oral cancer. Why betel quid components induce oral cancer is not clear. METHODS: Cytotoxicity of areca nut extract and arecoline (an areca nut alkaloid) to SAS oral epithelial cell line was evaluated by trypan blue dye exclusion and MTT assays. Cell cycle distribution and apoptosis was analyzed by propidium iodide staining flow cytometry. Chk1 and chk2 activation was analyzed by Pathscan phospho-enzyme-linked immunosorbent assay. Metalloproteinase-9 (MMP-9), tissue inhibitors of metalloproteinase (TIMPs) production was measured by enzyme-linked immunosorbent assay. RESULTS: Areca nut extract (800 µg/mL) and arecoline (>0.4 mmol/L) caused cell death, apoptosis, and cell cycle arrest of SAS cells. Areca nut extract and arecoline stimulated Chk1 and Chk2 phosphorylation in SAS cells. Areca nut extract stimulated cellular MMP-9 but suppressed TIMP-1 and TIMP-2 production. CONCLUSIONS: Areca nut components activate Chk1/Chk2, alter cell cycle regulation/apoptosis, MMP-9, and TIMPs production, contributing to the pathogenesis of oral carcinogenesis.

[Effects of phytohormones on hairy root growth and tanshinone biosynthesis of Salvia miltiorrhiza].

OBJECTIVE: To study the effects of different phytohormones on the growth of hairy root and biosynthesis of tanshinone II A in Salvia miltiorrhiza. METHODS: Salvia miltiorrhiza hairy root was induced by organizational culture method. Tanshinone was extracted with ultrasonic method and detected by high performance liquid chromatography (HPLC). RESULTS: The phytohormone combination of GA3 and 6-BA at different concentrations adding to 1/2 MS medium had obviously inhibitory effect on the hairy root growth of Salvia miltiorrhiza. And the highest inhibition rate was up to 100% (hairy roots were death). However, the growth of hairy roots could be obviously promoted by the combination of KT (1.0 mg/L) and IBA (1.0 mg/L), increasing 3. 251 times than that of the control. Total tanshinone II A content in hairy roots bodies was the highest under the combination of 0.2 mg/L NAA and 3.0 mg/L 6-BA. 6-BA of 2.0 mg/L played a significant role in promoting tanshinone II A biosynthesis, and the tanshinone II A concentration was 3.012 times higher than that of the control. CONCLUSION: Different phytohormones and its combination adding to 1/2 MS medium has significant influence on the hairy root growth and the content of tanshinone II A in Salvia miltiorrhiza.