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This study aims to explore the potential mechanism of Liangfu Pills in the treatment of functional dyspepsia(FD) based on network pharmacology and molecular docking, and verify the mechanism by animal experiment. The active components of Liangfu Pills were screened from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and the targets of Liangfu Pills were predicted by SwissTargetPrediction. The targets of FD were retrieved from GeneCards. On this basis, the common targets of the disease and the pills were yielded and the protein interaction was retrieved based on STRING. The core targets were screened out, followed by Gene Oncology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis with DAVID. Finally, molecular docking was carried out with the help of AutoDock Tools to predict the binding degree between the effective components of Liangfu Pills and core targets. A total of 19 active components of Liangfu Pills and 591 FD-related targets were screened out by network pharmacology, of which 253 were common targets of the disease and the prescription. Liangfu Pills was mainly involved in the biological processes of response to drug, negative regulation of transcription, positive regulation of apoptotic process, and cell surface receptor signaling pathway, and the KEGG pathways of hypoxia-inducible factor-1(HIF-1) signaling pathway, serotonergic synapse, tumor necrosis factor(TNF) signaling pathway, cyclic adenosine monophosphate(cAMP) signaling pathway, calcium signal pathway, and inflammatory mediator regulation of transient receptor potential(TRP) channels. The results of molecular docking showed that the key active components of Liangfu Pills had certain binding activity to the targets mitogen-activated protein kinase 1(MAPK1), protein kinase B(AKT1), transient receptor potential cation channel subfamily V member 1(TRPV1), 5-hydroxytryptamine receptor 1 A(HTR1 A), and 5-hydroxytryptamine receptor 2 A(HTR2 A). FD was induced in rats, and then Liangfu Pills was given to FD rats for 7 days. The results showed that Liangfu Pills could significantly relieve the symptoms of FD rats, significantly increase the expression of 5-hydroxytryptamine(5-HT), and down-regulate the expression of TRPV1. Through network pharmacology, molecular docking, and experimental verification, this study proved that Liangfu Pills improved FD through multiple components and multiple targets. The result lays a basis for further research on the mechanism and clinical application of Liangfu Pills in the treatment of FD.

Enterovirus 71 (EV71), a newly emerging life-threatening pathogen induces hand-foot-mouth disease (HFMD), no effective vaccines or specific anti-viral treatments are currently available. In this study, the activity of hederacolchiside C (HSC) against EV71 was investigated, and the antiviral mechanism was explored. HSC displayed apparent antiviral activity in EV71-infected cells probably through activating the host innate immunity. Comparing with EV71-infected group at 24 hpi, the group pretreated with HSC dramatically increased the expression of MAVS, p-IRF3, IRF3 and IFN-β, the innate immune effectors related to innate immunity. In addition, HSC displayed stronger antiviral activity in EV71-infected suckling mice in comparison with Ribavirin, a broad-spectrum antiviral drug. The results suggest that HSC could have potential as a pharmaceutical drug for HFMD.

In healthy joints, synovial fibroblasts (SFs) provide the microenvironment required to mediate homeostasis, but these cells adopt a pathological function in rheumatoid arthritis (RA). Carbohydrates (glycans) on cell surfaces are fundamental regulators of the interactions between stromal and immune cells, but little is known about the role of the SF glycome in joint inflammation. Here we study stromal guided pathophysiology by mapping SFs glycosylation pathways. Combining transcriptomic and glycomic analysis, we show that transformation of fibroblasts into pro-inflammatory cells is associated with glycan remodeling, a process that involves TNF-dependent inhibition of the glycosyltransferase ST6Gal1 and α2-6 sialylation. SF sialylation correlates with distinct functional subsets in murine experimental arthritis and remission stages in human RA. We propose that pro-inflammatory cytokines remodel the SF-glycome, converting the synovium into an under-sialylated and highly pro-inflammatory microenvironment. These results highlight the importance of glycosylation in stromal immunology and joint inflammation., Dysregulation of synovial fibroblasts is thought to be an important step in the pathogenesis of rheumatoid arthritis. Here the authors implicate α2-6 sialylation in this process by studying the glycome of these cells in patients and in a mouse model of inflammatory joint disease.

The progressive increase of metabolic diseases underscores the necessity for developing effective therapies. Although we found Tian-Huang formula (THF) could alleviate metabolic disorders, the underlying mechanism remains to be fully understood. In the present study, firstly, male Sprague-Dawley rats were fed with high-fat diet plus high-fructose drink (HFF, the diet is about 60% of calories from fat and the drink is 12.5% fructose solution) for 14 weeks to induce hepatosteatosis and glucose intolerance and then treated with THF (200 mg/kg) for 4 weeks. Then, metabolomics analysis was performed with rat liver samples and following the clues illustrated by Ingenuity Pathway Analysis (IPA) with the metabolomics discoveries, RT-qPCR and Western blotting were carried out to validate the putative pathways. Our results showed that THF treatment reduced the body weight from 735.1 ± 81.29 to 616.3 ± 52.81 g and plasma triglyceride from 1.5 ± 0.42 to 0.88 ± 0.33 mmol/L; meanwhile, histological examinations of hepatic tissue and epididymis adipose tissue showed obvious alleviation. Compared with the HFF group, the fasting serum insulin and blood glucose level of the THF group were improved from 20.77 ± 6.58 to 9.65 ± 5.48 mIU/L and from 8.96 ± 0.56 to 7.66 ± 1.25 mmol/L, respectively, so did the serum aspartate aminotransferase, insulin resistance index, and oral glucose tolerance ( p = 0.0019, 0.0053, and 0.0066, respectively). Furthermore, based on a list of 32 key differential endogenous metabolites, the molecular networks generated by IPA suggested that THF alleviated glucose intolerance and hepatosteatosis by activating phosphatidylinositol-3 kinase (PI3K) and low-density lipoprotein receptor (LDL-R) involved pathways. RT-qPCR and Western blotting results confirmed that THF alleviated hepatic steatosis and glucose intolerance partly through protein kinase B- (AKT-) sterol regulatory element-binding protein (SREBP) nexus. Our findings shed light on molecular mechanisms of THF on alleviating metabolic diseases and provided further evidence for developing its therapeutic potential.

Background & Aims Nonalcoholic fatty liver disease (NAFLD) has become the most common chronic liver diseases worldwide. The advantage stage of NAFLD, nonalcoholic steatohepatitis (NASH), has been recognized as the leading cause of end-stage liver injury without FDA-approved therapeutic strategy. Glutathione S-transferase Mu 2 (GSTM2) is a phase II detoxification enzyme. However, the roles of GSTM2 in NASH has not been elucidated yet. Methods Multiple RNA-seq analyses were used to identify hepatic GSTM2 expression in NASH. In vitro and in vivo gain- or loss-of-function approaches were used to investigate the role and molecular mechanism of GSTM2 in NASH. Results We identified glutathione S-transferase Mu 2 (GSTM2) as a sensitive responder and effective suppressor of NASH progression. GSTM2 was significantly downregulated during NASH progression. Hepatocyte GSTM2 deficiency markedly aggravated insulin resistance, hepatic steatosis, inflammation and fibrosis induced by a high-fat diet and a high-fat/high-cholesterol diet. Mechanistically, GSTM2 sustained MAPK pathway by directly interacting with apoptosis signal-regulating kinase 1 (ASK1). Furthermore, GSTM2 directly bound to the amino-terminal (N-terminal) region of ASK1 and inhibited ASK1 N-terminal dimerization to subsequently repress ASK1 phosphorylation and the activation of its downstream JNK/p38 signaling pathway under metabolic dysfunctional condition. Conclusions These data demonstrated that hepatocyte GSTM2 was an endogenous suppressor protecting against NASH progression by blocking ASK1 N-terminal dimerization and phosphorylation. This study paved a new way for the development of therapeutic strategy for NASH by activating GSTM2. Lay summary The GSTM2 has a major role in regulating NASH development. We identified that GSTM2 could exert beneficial effects as an endogenous suppressor of ASK1 dimerization and its downstream ASK1-JNK/p38 signaling activation. Targeting GSTM2 is a promising therapeutic strategy for the treatment of NASH. Clinical trial number IIT-2021-277

In healthy joints, synovial fibroblasts (SFs) provide the microenvironment required to mediate homeostasis but are recognized to adopt a pathological role in rheumatoid arthritis (RA), promoting the infiltration and activation of immune cells to perpetuate local inflammation, pain and joint destruction. Carbohydrates (glycans) attached to cell surface proteins are fundamental regulators of cellular interactions between stromal and immune cells, but very little is known about the glycome of SFs or how glycosylation regulates their biology. Here we fill these gaps in our understanding of stromal guided pathophysiology by systematically mapping glycosylation pathways in healthy and arthritic SFs. We used a combination of transcriptomic and glycomic analysis to show that transformation of fibroblasts into pro-inflammatory cells in RA is associated with profound glycan remodeling, a process that involves reduction of α2-6 terminal sialylation that is mostly mediated by TNFα-dependent inhibition of the glycosyltransferase ST6Gal1. We also show that sialylation of SFs correlates with distinct disease stages and SFs functional subsets in both human RA and models of mouse arthritis. We propose that pro-inflammatory cytokines in the joint remodel the SF-glycome, transforming a regulatory tissue intended to preserve local homeostasis, into an under-sialylated and highly pro-inflammatory microenvironment that contributes to an amplificatory inflammatory network that perpetuates chronic inflammation. These results highlight the importance of cell glycosylation in stromal immunology.

Among different types, Chinese propolis (ChPs) and Brazilian green propolis (BrGPs) have been shown to contain multi-functional properties. Despite extensive research in the field, reports comparing propolis from different geographical areas are still limited, compromising our current understanding of the potential therapeutic effect associated with propolis and its derived compounds. Herein, a comparative study between ChPs and BrGPs including their metabolite profile and bioactivities was performed. Interestingly, even when ChPs and BrGPs showed similar anti-inflammatory potential, our results showed that they contained very different levels of ethanol extract, total flavonoids and total phenolic acids and in fact, LC-MS metabolic profiling and pattern recognition could effectively distinguish ChPs and BrGPs. Moreover, all the propolis samples tested showed good anti-oxidant activity and no significant difference of free radical scavenging capacity existed between ChPs and BrGPs. In conclusion, ChPs and BrGPs have a distinct chemome, but their antioxidant and anti-inflammatory activities are similar.

Microctis Folium (MF), the dried leaves of Microcos paniculata, is widely used as a medical and food dual-purpose herb in South-east Asia and China. However, the quality control of MF is not well studied. A simple and reliable quality control method was urgently needed for its growing usage. Herein, at first, its main active components were identified by UPLC-ESI-qTOF-MS, and a representative MF flavone glycosides profile consisting of ten compounds was illustrated, which is the most detailed one up to now. Successively, using vitexin as the reference substance, a novel QAMS method with HPLC for quantification of the ten identified flavone glycosides was developed and methodologically validated. Furthermore, making use of the abovementioned QAMS method, quantitative profiling of 21 batches of prepared MF slices collected from different hospital pharmacies were performed. As a result, the total contents of ten flavone glucosides and the content of specific compound showed obvious variations. Using the ten compounds’ contents dataset, the 21 batches of samples were divided into two distinct clusters by HCA. In sum, our results indicated that it was of great importance to take quality control of prepared MF slices and we presented a robust and simple method for their quantitative determination, which should be beneficial for the quality control of MF and its derived products.

Desmodium styracifolium is one of the traditional Chinese herbs. In the present study, membrane-based technologies were used to separate and concentrate of the total flavone glycosides fraction from D. styracifolium. The extracts flowed through an ultrafiltration membrane which MWCO (molecular weight cut-off) is 30KDa and the permeate was concentrated by MWCO-1KDa nano-filtration membrane. The solid content of the membrane concentrated extracts of D. styracifolium (MEDs) was 26.5 mg/ml. Moreover, the content of vicenin-2, schaftoside and isovitexin in MEDs were 4.88 %, 9.76 %, 1.89 % respectively. The assay in vitro showed MEDs has better anti-inflammatory effect which partly proved that our membrane-based processes for separation and concentration of flavone glycosides from D. styracifolium is reliable and practicable.

Ample evidence supports the prominent role of gut-liver axis in perpetuating pathological networks of high-fat high-fructose (HFF) diet induced metabolic disorders, however, the molecular mechanisms are still not fully understood. Herein, this study aims to present a holistic delineation and scientific explanation for the crosstalk between the gut and liver, including the potential mediators involved in orchestrating the metabolic and immune systems.An experimental obesity-associated metaflammation rat model is induced with a HFF diet. An integrative multi-omics analysis is then performed. Following the clues illustrated by the multi-omics discoveries, putative pathways are subsequently validated by RT-qPCR and Western blotting. HFF diet leads to obese phenotypes in rats, as well as histopathological changes. Integrated omics analysis shows that there exists a strong interdependence among gut microbiota composition, intestinal metabolites, and innate immunity regulation in the liver. Some carboxylic acids may contribute to gut-liver communication. Moreover, activation of the hepatic LPS-TLR4 pathway in obesity is confirmed.HFF-intake disturbs gut flora homeostasis. Crosstalk between gut microbiota and innate immune system mediates hepatic metaflammation in obese rats, associated with LPS-TLR4 signaling pathway activation. Moreover, α-hydroxyisobutyric acid and some other organic acids may play a role as messengers in the liver-gut axis.

Acute lung injury (ALI) and its more severe form acute respiratory distress syndrome (ARDS) are life-threatening conditions with high morbility and mortality, underscoring the urgent need for novel treatments. Leaves of the medicinal herb Microcos paniculata have been traditionally used for treating upper airway infections, by virtue of its content of flavonoids such as apigenin C-glycosides (ACGs). C-glycosides have been shown to exert strong anti-inflammatory properties, although their mechanism of action remains unknown. Herein, hypothesizing that ACGs from M. paniculata inhibit progression of ALI, we used the experimental model of lipopolysaccharide (LPS)-induced ALI in BALB/c mice to evaluate the therapeutic potential of purified ACGs. Our results showed that M. paniculata ACGs inhibited lung inflammation in animals undergoing ALI. The protective effects of ACGs were assessed by determination of cytokine levels and in situ analysis of lung inflammation. ACGs reduced the pulmonary edema and microvascular permeability, demonstrating a dose-dependent down-regulation of LPS-induced TNF-α, IL-6 and IL-1β expression in lung tissue and bronchoalveolar lavage fluid, along with reduced apoptosis. Moreover, metabolic profiling of mice serum and subsequent Ingenuity Pathway Analysis suggested that ACGs activated protective protein networks and pathways involving inflammatory regulators and apoptosis-related factors, such as JNK, ERK1/2 and caspase-3/7, suggesting that ACGs-dependent effects were related to MAPKs and mitochondrial apoptosis pathways. These results were further supported by evaluation of protein expression, showing that ACGs blocked LPS-activated phosphorylation of p38, ERK1/2 and JNK on the MAPKs signaling, and significantly upregulated the expression of Bcl-2 whilst down-regulated Bax and cleaved caspase-3. Remarkably, ACGs inhibited the LPS-dependent TLR4 and TRPC6 upregulation observed during ALI. Our study shows for the first time that ACGs inhibit acute inflammation and apoptosis by suppressing activation of TLR4/TRPC6 signaling pathway in a murine model of ALI. Our findings provide new evidence for better understanding the anti-inflammatory effects of ACGs. In this regard, ACGs could be exploited in the development of novel therapeutics for ALI and ARDS.

Objective Hederacolchiside A1, exhibits cytostatic and cytotoxic activity against various cancer cells in vitro, however, the mechanism is not well understood. Methods In this study, Hederacolchiside A1 from Pulsatilla chinensis was isolated and tested its anticancer activity and mechanism. Hederacolchiside A1 could inhibit proliferation of A549, SMMC-7721, BEL-7402, and MCF-7 cells by MTT assay. Investigations of apoptosis of treated cancer cells were identified in hederacolchiside A1 by flow cytometric analysis of annexin V expression. Results Based on the results of western blotting and JC-1 staining, hederacolchiside A1 reduced the mitochondrial membrane potential and Bcl-2 protein levels, whereas cleaved caspase-3 was higher. Furthermore, hederacolchiside A1 effectively inhibited the phosphorylations of phosphatidylinositol 3 kinase (PI3K), protein kinase B (Akt), and mammalian target of rapamycin (mTOR). In vivo study showed that hederacolchiside A1 (3.0, 4.5, and 6.0 mg/kg, ip) could significantly inhibit the weight of tumor in an H22 xenograft model. Similar inhibitory activities were observed when the compound (3.25, 7.5, and 15.0 mg/kg, ig) was tested in nude mice xenograft tumor models using human breast carcinoma MCF-7 cells. Conclusion These data indicated that hederacolchiside A1 suppressed the proliferation of human tumor cells by inducing apoptosis through modulating the PI3K/Akt/mTOR signaling pathway.

Scope Considerable evidence supports the view that high-fructose intake is associated with increased and early incidence of obesity and dyslipidemia. However, knowledge on physiopathological alterations introduced by fructose overconsumption is lacking. Therefore, an integrated omics analysis is carried out to investigate the consequences of short-term fructose overfeeding (SFO) and identify the underlying molecular mechanisms. Methods and results SFO of rats demonstrates obvious histopathological hepatic lipid accumulation and significant elevation in adiposity, total cholesterol, and fasting plasma glucose levels. Integrated omics analysis demonstrates that SFO disturbed metabolic homeostasis and initiated metabolic stress. Hepatic lipogenesis pathways are also negatively impacted by SFO. Analysis of molecular networks generated by ingenuity pathway analysis (IPA) implicates involvement of the extracellular signal regulated kinase (ERK) signaling pathway in SFO and its consequences. Moreover, it is identified that an inherent negative feedback regulation of hepatic sterol regulatory element binding protein 1 (SREBP1) plays an active role in regulating hepatic de novo lipogenesis. Conclusion The findings indicate that SFO disturbs metabolic homeostasis and that endogenous small molecules positively mediate SFO-induced metabolic adaption. The results also underline that an inherent regulatory mechanism of resilience occurs in response to fructose overconsumption, suggesting that efforts to maintain resilience can be a promising target to prevent and treat metabolic disorder-like conditions.

Colorectal cancer (CRC) remains one of the most common cancers worldwide. Increasing evidence indicates that SPRY4 intronic transcript 1 (SPRY4-IT1) regulate cell growth, differentiation, apoptosis, and cancer progression. However, the expression and function of SPRY4-IT1 in the progression of CRC remains largely unknown. Here, we reported that SPRY4-IT1 was upregulated in CRC. Increased SPRY4-IT1 expression in CRC was associated with larger tumor size and higher clinical stage. In vitro experiments revealed that SPRY4-IT1 knockdown significantly inhibited CRC cell proliferation by causing G1 arrest and promoting apoptosis, whereas SPRY4-IT1 overexpression promoted cell proliferation. Further functional assays indicated that SPRY4-IT1 overexpression significantly promoted cell migration and invasion by regulate the epithelial-mesenchymal transition (EMT). Taken together, our study demonstrates that SPRY4-IT1 could act as a functional oncogene in CRC, as well as a potential therapeutic target to inhibit CRC metastasis.

Colorectal cancer (CRC) is one of the most common types of cancer worldwide. Recently, microRNAs (miRs) have been considered as novel therapeutic targets for the treatment of cancer. miR‑598 is a poorly investigated miR. The underlying mechanism of miR‑598 in CRC cells remains to be elucidated. In the present study, miR‑598 was demonstrated to be significantly upregulated in CRC tissue by analyzing data from The Cancer Genome Atlas and the Gene Expression Omnibus. The results of a polymerase chain reaction demonstrated that miR‑598 expression was significantly upregulated in CRC tissues and cells. Gain of function and loss of function assays demonstrated that miR‑598 significantly promoted cell proliferation and cell cycle progression. miR‑598 was demonstrated to modulate cell functions by regulating 72 kDa inositol polyphosphate‑5‑phosphatase (INPP5E). In addition, knockdown of INPP5E counteracted the growth arrest caused by an miR‑598‑inhibitor. In conclusion, the present study demonstrated that miR‑598 contributed to cell proliferation and cell cycle progression in CRC by targeting INPP5E.

Osteopontin (OPN) plays an important role in the metastasis and recurrence of tumors after resection of hepatocellular carcinoma (HCC). In this study, the down-regulation effect on OPN expression in HCC cells of RNA interference (RNAi) molecules designed to target different segments of OPN was investigated to identify a more effective site for OPN knockdown. Specific small interfering RNAs (siRNAs A, B, and C) of OPN were synthesized and transfected into an HCC cell line (HEP-G2; representing the OPNi-A, OPNi-B, and OPNi-C groups). Fluorescent quantitative polymerase chain reaction and immunohistochemical methods were used to detect the mRNA and protein expression of OPN before and after RNAi. Results showed that after transfection, the fluorescence intensity of the OPNi-A group was greater than those of the OPNi-B and OPNi-C groups. After 48 h of transfection, the ΔCT values of OPN mRNA expression in the OPNi-A-C groups increased from 8.31 ± 1.58, 8.78 ± 1.49, and 8.25 ± 1.51 to 12.14 ± 1.43, 10.22 ± 1.97, and 10.48 ± 1.88, respectively (P < 0.05), and the OPN protein levels (immunohistochemistry scores) decreased from 6.44 ± 1.67, 5.43 ± 2.05, and 5.45 ± 2.52 to 2.84 ± 1.52, 4.43 ± 1.65, and 3.95 ± 1.43 points, respectively. These results indicated that RNAi based on different segments of the OPN gene had different down-regulatory effects on OPN expression. Synthesis of targeted siRNA aimed at specific OPN segments might have important significance for dealing with the invasiveness and metastasis of HCC cells.

Microcos paniculata is an edible and medicinal plant in Lingnan, China. The present study investigated the anti-inflammatory effect and chemical constituents of its’ total flavone glycosides (MpTFG) fraction. MpTFG (10, 15, 20 μg/mL) was applied to 1 μg/mL LPS-induced RAW 264.7 macrophages in vitro; and MpTFG (10, 20, 40 mg/kg body weight) was employed to xylene-induced ear edema test. As a result, 20 μg/mL MpTFG decreased the levels of pro-inflammatory cytokines IL-6, IL-1β and TNF-α in LPS-stimulated RAW 264.7 cells supernatant significantly, and 40 mg/kg MpTFG could effectively ameliorate xylene-induced mice ear swelling. The results showed that MpTFG had potential anti-inflammatory effect. Meanwhile, ten compounds were purified by making use of macro-porous resin, sephadex LH-20 gel column chromatography and semi-preparation HPLC successively. They were characterized to be vicenin-2 (i), isoschaftoside (ii), schaftoside (iii), vitexin (iv), vicenin-1 (v), isovitexin (vi), isoviolanthin (vii), nicotiflorin (viii), astragalin (ix), narcissoside (x) by 1H-NMR, 13C-NMR and HRMS spectral analysis.

In order to improve the quality control method for propolis, HPLC fingerprinting and pattern recognition of 12 batches of Brazilian green propolis and 12 batches of Chinese propolis were carried out. HPLC fingerprinting was performed on a C18 column (250 mm × 4.6 mm, 5μm) using gradient elution with methanol-water (containing 0.2% formic acid) as the mobile phase. Similarity analysis was executed on similarity evaluation system for chromatographic fingerprint of TCM (2012A). Pattern recognition was performed by SIMCA-P 13.0 software. As a result, as for the two groups of propolis samples, 21 common peaks were calibrated and 11 of them were identified as chlorogenic acid, caffeic acid, isochlorogenic acid B, isochlorogenic acid C, quercetin, kaempferol, apigenin, pinocembrin, caffeic acid phenylethyl ester, galangin and artepillin C. The similarity of the fingerprint profiles of the tested Brazilian propolis is more than 0.92, and that of the tested Chinese propolis is more than 0.91. However, the two groups of fingerprints’ similarity are from 0.523 to 0.693. The results of PCA and OPLS-DA showed that chemical profiles of the tested Brazilian green propolis were different from those of Chinese propolis. A set of compounds, namely, artepillin C, chlorogenic acid, pinocembrin, isochlorogenic acid B and caffeic acid phenylethyl ester can act as difference marker of the two groups of propolis. Moreover, HPLC fingerprint combined with pattern recognition could be developed into one new method for quality control of propolis.

Brazilian green propolis is a complex mixture of natural compounds that is difficult to analyze and standardize, as a result, controlling its quality is challenging. In this study, we used the positive and negative modes of ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time of flight mass spectrometry in conjunction with high-performance liquid chromatography for the identification and characterization of seven phenolic acid compounds in Brazilian green propolis. The optimal operating conditions for the electrospray ionization source were capillary voltage of 3500 V and drying and sheath gas temperatures of 320 &deg, C and 350 &deg, C, respectively. Drying and sheath gas flows were set to 8 L/min and 11 L/min, respectively. Brazilian green propolis was separated using the HPLC method, with chromatograms for samples and standards measured at 310 nm. UPLC-ESI-QTOF-MS was used to identify the following phenolic compounds: Chlorogenic acid, caffeic acid, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C, caffeic acid phenethyl ester (CAPE), and artepillin C. Using a methodologically validated HPLC method, the seven identified phenolic acids were then quantified among different Brazilian green propolis. Results indicated that there were no significant differences in the content of a given phenolic acid across different Brazilian green propolis samples, owing to the same plant resin sources for each sample. Isochlorogenic acid B had the lowest content (0.08 &plusmn, 0.04) across all tested Brazilian green propolis samples, while the artepillin C levels were the highest (2.48 &plusmn, 0.94). The total phenolic acid content across Brazilian green propolis samples ranged from 2.14&ndash, 9.32%. Notably, artepillin C quantification is an important factor in determining the quality index of Brazilian green propolis, importantly, it has potential as a chemical marker for the development of better quality control methods for Brazilian green propolis.

Two new 28-nor-oleanane-type triterpene saponins, oleiferoside U (1), and oleiferoside V (2) were isolated from the 50% EtOH extract of the roots of Camellia oleifera C. Abel. Their structures were elucidated as camellenodiol 3β-O-β-d-galactopyranosyl-(1→2)-β-d-xylopyranosyl-(1→2)-[β-d-galactopyranosyl-(1→3)]-β-d-glucuronopyranoside and camellenodiol 3β-O-β-d-galactopyranosyl-(1→3)-β-d-xylopyranosyl-(1→2)-[β-d-galactopyranosyl-(1→3)]-β-d-glucuronopyranoside. Their chemical structures were established mainly on the basis of integrated spectroscopic techniques. In vitro, cytotoxic activities of the two new triterpene saponins were evaluated against three human tumor cell lines (A549, SMMC-7721, and MCF-7) using the MTT assay. Both of them showed a certain cytotoxic activities toward the tested cell lines and gave IC50 values in the range of 45.04-63.22 μM.

This study investigated the clinical pathologic character of malignant gastrointestinal stromal tumors (MGIST), their treatment with surgery, and evaluated the efficacy of imatinib postoperation. A total of 68 MGIST patients were enrolled. Of these, 27 patients underwent imatinib auxiliary therapy (treatment group) and 41 underwent imatinib therapy (control group). The therapeutic effects on the two groups were compared using χ2 test analysis after follow-up of two years. The expressions of CD117, CD34, S100, Vimentin, and alpha smooth-muscle actin (SMA) were detected by immunohistochemistry methods. Of the 68 cases, 28 showed potential MGIST, whereas 40 had MGIST. Haemorrhagia or necrosis, abundant cell, manifest heteromorphism, and caryocinesia were observed in varying degrees. The positive rates of CD117, CD34, Vimentin, S100, and SMA were 89.7% (61/62), 88.2% (60/62), 73.5% (50/62), 41.1% (28/62) and 25.0% (17/62), respectively. The recurrence rate in the treatment group was significantly lower than that in the control group (p

To establish a new method for the extraction and separation of curcuminoids from Curcuma longa rhizome by cloud-point preconcentration using microemulsions as solvent.The spectrophotometry was used to detect the solubility of curcumin in different oil phase, emulsifier and auxiliary emulsifier, and the microemulsion prescription was used for false three-phase figure optimization. The extraction process was optimized by uniform experiment design. The curcuminoids were separated from microemulsion extract by cloud-point preconcentration.Oil phase was oleic acid ethyl ester; Emulsifier was OP emulsifier; Auxiliary emulsifier was polyethylene glycol(peg) 400; The quantity of emulsifier to auxiliary emulsifier was the ratio of 5: 1; Microemulsion prescription was water-oleic acid ethyl ester-mixed emulsifier (0.45:0.1:0.45). The optimum extraction process was: time for 12.5 min, temperature of 52 degrees C, power of 360 W, frequency of 400 kHz, and the liquid-solid ratio of 40:1. The extraction rate of curcuminoids was 92.17% and 86.85% in microemulsion and oil phase, respectively.Curcuminoids is soluble in this microemulsion prescription with good extraction rate. This method is simple and suitable for curcuminoids extraction from Curcuma longa rhizome.

High recurrence of colon cancer liver metastasis is observed in patients after hepatic surgery, and the cause is believed to be mostly due to the growth of residual microscopic metastatic lesions within the residual liver. Therefore, triggering the progression of occult metastatic foci may be a novel strategy for improving survival from colon cancer liver metastases. In the present study, we identified an anti-recurrence effect of ulinastatin on colon cancer liver metastasis in mice after hepatectomy. Transwell cell invasion assays demonstrated that ulinastatin significantly inhibited thein vitroinvasive ability of colon cancer HCT116 cells. Moreover, gelatin zymography and ELISA analysis showed that MMP-9 activity and plasmin activity of colon cancer HCT116 cells were inhibited by ulinastatin, respectively. Furthermore,in vivoBALB/C nu/nu mice model indicated that ulinastatin effectively reduced recurrence after resection of hepatic metastases from colon cancer. The optimum timing for ulinastatin administration was one week after hepatectomy. Taken together, our findings point to the potential of ulinastatin as an effective approach in controlling recurrence of hepatic metastases from colon cancer after hepatectomy via its anti-plasmin activity.

The aim of the present study was to investigate the inhibitory effects of thalidomide in the hepatocellular carcinoma nude mouse model in order to provide new insights into a comprehensive clinical intervention for hepatocellular carcinoma. MHCC97 cells were routinely cultured, passaged and adjusted to a single cell suspension with a concentration of 2×107/ml. Six-week-old, BALB/C male nude mice were anesthetized and fixed in the prone position, then a subcapsular injection of the single cell suspension was administered into the spleen and their abdomens were closed. A laparotomy and left hepatic lobectomy was performed 14 days later and the abdomens were closed once again. Subsequent to the establishment of the hepatocellular carcinoma model, the nude mice were randomly divided into three groups, each consisting of 12 mice. The early intervention group were immediately provided with the post-operative thalidomide intervention, the late intervention group were provided with the post-operative thalidomide intervention one week subsequent to the surgery, and the negative control group were provided with a placebo intervention (0.9% physiological saline). Each intervention was continuously administered once per day for one week. The osteopontin (OPN) content of the liver tumors was detected using immunohistochemistry. The data were analyzed using an analysis of variance (ANOVA) test. There were significant differences in the OPN levels of the tumors among the early intervention, late intervention and negative control groups. Thalidomide may inhibit the generation of OPN and thereby inhibit the infiltration and metastasis of tumors; the immediate use of thalidomide following hepatectomy in the present study may block the invasion and metasis for liver cancer more effectively.

To establish an optimum enrichment and purification process of total flavonoids in Microcos paniculata by macroporous resins.Five kinds of resins were compared and the best one was chosen. Then the parameters of the process were optimized by single factor tests, uniform design and statistical methods.DI01 was selected for its excellent adsorption and desorption properties, 70% ethanol was found to be the best elution solution. As far as the yield was considered, the best result was based on the followings: feed rate-1.0 BV/h, elution flow rate-2.0 BV/h, sample concentration-7.88 mg/mL, eluting agent amount-2.0 BV, pH value 4.8; then the yield reached 90.18% and the purity was 54.37%. If the purity was considered, the best parameters wereas follows: feed rate-1.0 BV/h, elution flow rate-2.0 BV/h, sample concentration-2.0 mg/mL, eluting agent amount-2.8 BV, pH value 7.8; then the purity reached 61.77% and the yield was 80.25%.The total flavonoids of Microcos paniculata can be effectively purificated and separated by D101 macroporous resin.