Methyl jasmonate regulation of pectin polysaccharides in Cosmos bipinnatus roots: A mechanistic insight into alleviating cadmium toxicity.

Methyl jasmonate (MeJA), a crucial phytohormone, which plays an important role in resistance to Cadmium (Cd) stress. The cell wall (CW) of root system is the main location of Cd and plays a key role in resistance to Cd toxicity. However, the mechanism effect of MeJA on the CW composition and Cd accumulation remain unclear. In this study, the contribution of MeJA in regulating CW structure, pectin composition and Cd accumulation was investigated in Cosmos bipinnatus. Phenotypic results affirm MeJA's significant role in reducing Cd-induced toxicity in C. bipinnatus. Notably, MeJA exerts a dual impact, reducing Cd uptake in roots while increasing Cd accumulation in the CW, particularly bound to pectin. The molecular structure of pectin, mainly uronic acid (UA), correlates positively with Cd content, consistent in HC1 and cellulose, emphasizing UA as pivotal for Cd binding. Furthermore, MeJA modulates pectin methylesterase (PME) activity under Cd stress, influencing pectin's molecular structure and homogalacturonan (HG) content affecting Cd-binding capacity. Chelate-soluble pectin (CSP) within soluble pectins accumulates a substantial Cd proportion, with MeJA regulating both UA content and the minor component 3-deoxy-oct-2-ulosonic acid (Kdo) in CSP. The study delves into the intricate regulation of pectin monosaccharide composition under Cd stress, revealing insights into the CW's physical defense and Cd binding. In summary, this research provides novel insights into MeJA-specific mechanisms alleviating Cd toxicity in C. bipinnatus, shedding light on complex interactions between MeJA, and Cd accumulation in CW pectin polysaccharide.

Explore the pharmacological basis of ShengJiYiSui decoction in the treatment of amyotrophic lateral sclerosis based on network pharmacology and molecular docking technology.

Neurodegenerative illnesses have long been handled clinically by traditional Chinese medicine. This study is the first time to explore the pharmacological basis of application in amyotrophic lateral sclerosis (ALS) through network pharmacology and molecular docking techniques. In the present investigation, the TCMSP database and HIT2 database were examined for 9 TCM constituents of Sheng Ji Yu Sui Decoction (SJYSD), and the desired sites for the components were searched in the Drugbank database. and the Sjysd-target network was constructed. Associated targets for Amyotrophic lateral sclerosis (ALS) were then retrieved and collected in the OMIM, TTD, Genecards and DisGeNET databases. Protein-protein interaction and enrichment analysis were performed for the common targets of drugs and diseases, and molecular anchoring for the chosen core targets and related molecules was carried out. The results showed that SJYSD had 100 active compounds corresponding to 598 targets. ALS has a total of 5,325 genes. SJYSD and ALS share 163 genes, and these targets involve PI3K-AKT signaling, p53 signaling and IL-17 signaling, etc. The core components of luteolin and quercetin were discovered and may be used to treat ALS by regulating PI3K-AKT signaling pathway by HSP90AB1 protein.

Effects of nitrogen and phosphorus supply levels and ratios on soil microbial diversity-ecosystem multifunctionality relationships in a coastal nontidal wetland.

Human activities have changed the levels and ratios of nitrogen (N) and phosphorus (P) in wetland ecosystems. However, the effects of N and P levels and ratios on wetland soil microbial community and ecosystem multifunctionality remain unclear, especially on the relationships between soil microbial diversity and ecosystem multifunctionality. In this study, the effects of a 7-year experimental nutrient addition on the soil microbial community and ecosystem multifunctionality (12 function variables related to carbon, N, and P cycling) were assessed by combining three N and P supply levels with three N:P supply ratios in a coastal nontidal wetland ecosystem. According to the obtained results, the N and P supply levels significantly affected soil bacterial community composition, as well as ecosystem multifunctionality, while no significant effects of N:P supply ratios were observed. Although N and P supply levels did not significantly affect bacterial and fungal diversity, they both changed the complexity of bacterial and fungal networks. Soil ecosystem multifunctionality was significantly and positively correlated with bacterial diversity rather than fungal diversity. Moreover, the correlation coefficient between bacterial diversity and ecosystem multifunctionality showed an increasing-decreasing trend with increasing N and P supply levels and an increasing trend with increasing N:P supply ratios. However, the correlation coefficient between bacterial diversity and ecosystem multifunctionality was not significantly correlated with bacterial network complexity. The current study provides new insights into the impacts of N and P levels and ratios on soil microbial community and ecosystem multifunctionality in a coastal nontidal wetland. In particular, the present study highlighted that changes in N and P supply levels and ratios lead to changes in the relationship between soil bacterial diversity and ecosystem multifunctionality, which should be considered in related studies to accurately predict the responses of ecosystem multifunctionality to N and P inputs in coastal nontidal wetlands.

Direct electrochemical detection of Cu(â ¡) ions in juice and tea beverage samples using MWCNTs-BMIMPF6-Nafion modified GCE electrodes.

Due to a small amount of Cu (Ⅱ) ions being beneficial and too much being harmful, it is necessary to establish a rapid and direct detection method. Herein, we reported a platform based on multiwalled carbon nanotubes (MWCNTs), 1-butyl-3-methylimidazolium hexafluorophosphate (BMIMPF6), and Nafion solution-modified glassy carbon electrode (GCE) for the direct electrochemical detection of Cu (II) ions. We used differential pulse anodic stripping voltammetry, including the electrodeposition of Cu (Ⅱ) ions on the modified GCE and subsequent anodic stripping. Under the optimum conditions, the linear range was 20 µg·L-1 âˆ¼ 950 µg·L-1, the limit of detection (LOD) was 16 µg·L-1, and the limit of quantification (LOQ) was 54 µg·L-1 for Cu (II). We realized the quantitative detection of Cu (Ⅱ) ions in juice and tea beverage without tedious pretreatment. The result showed that the sensor had good anti-interference and practicability for actual food samples.

Effect of baotaiyin on IL-23 /Th17 immune inflammatory axis in mouse model of spontaneous abortion. / ä¸­è¯ä¿èƒŽé¥®å¯¹è‡ªç„¶æµäº§å°é¼ æ¨¡åž‹IL-23/Th17å ç–«ç‚Žç—‡è½´çš„å½±å“.

OBJECTIVES: The mechanism for traditional Chinese medicine in treating of recurrent spontaneous abortion is not clear. This study aims to explore the mechanism of baotaiyin in the treatment of recurrent abortion by regulating the immune inflammatory axis of interleukin (IL)-23/helper T cell (Th)17. METHODS: Spontaneous abortion model mice were randomly divided into a model group, 3 dose (low, medium, and high) groups of baotaiyin, with 10 mice in each group. After 14 days of medication, the levels of IL-17, IL-23, IL-10, and TGF-ß in serum were detected with enzyme-linked immunosorbent assay. The proportion of Th17 and regulatory T cells (Treg) cells in spleen lymphocytes was tested with flow cytometry. The expressions of (retinoid-related orphan receptor γt, ROR-γt) and forkhead box P3 (FOXP3) mRNA in decidua tissues was detected with RT-PCR. Embryo absorption rate was counted. RESULTS: Compared with the model group, the absorption rate of embryo and Th17/Treg cell ratio in baotaiyin medium- and high-dose groups were decreased significantly (all P<0.05); the levels of IL-17 and IL-23 in serum were decreased (both P<0.05), while the levels of TGF-ß and IL-10 in baotaiyin medium- and high-dose groups were increased (P<0.05, P<0.01, respectively); the expression of ROR-γt mRNA was decreased and the expression of FOXP3 mRNA was increased (all P<0.01) in decidua tissues of baotaiyin medium- and high-dose groups. CONCLUSIONS: Baotaiyin inhibits the positive feedback cycle of IL-23/Th17 immune inflammatory axis, which regulates Th17/Treg cell balance, mediates the maternal and fetal immune tolerance, and prevents the recurrent abortion.

Effect of electro-acupuncture on ovarian function of women with diminished ovarian reserve: study protocol for a randomized controlled trial.

BACKGROUND: Diminished ovarian reserve (DOR) is the precursor state of ovarian failure and can cause the decline of women's reproductive function. DOR also leads to poor outcome of in vitro fertilization and embryo transfer (IVF-ET) by affecting the oocytes, high qualified embryo rate, pregnancy rate, etc. Some studies have demonstrated that acupuncture can improve ovarian function. But to date, there is limited evidence indicating that acupuncture or electro-acupuncture is efficient to DOR. This trial aims to evaluate the efficiency and safety of electro-acupuncture for the ovarian function and the following outcome of IVF-ET in DOR patients. METHODS: This will be a multicenter randomized controlled clinical trial. A total of more than 338 women with DOR will be randomly allocated to treatment and control groups in 1:1 ratio receiving acupuncture before undergoing IVF-ET. The primary outcome will be the clinical pregnancy rate per cycle of IVF-ET after acupuncture. The secondary outcomes will be ovarian reserve function, outcomes of IVT-ET, blood biochemical index, Massachusetts General Hospital Acupuncture Sensation Scale (MASS), scores from the self-rating anxiety and depression scale, quality of life, and pregnancy outcomes. The safety of acupuncture will also be assessed. DISCUSSION: The results of this trial may provide high-quality evidence regarding the effectiveness of electro-acupuncture in the treatment of DOR and following outcomes of IVF-ET. This will also help patients with DOR and their physicians by offering a new treatment option. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR1900024626 . Registered on 19 July 2019.

Highly Crystalline Covalent Organic Frameworks Act as a Dual-Functional Fluorescent-Sensing Platform for Myricetin and Water, and Adsorbents for Myricetin.

Selective detection of active ingredients in complex samples has always been a crucial challenge because there are many disturbing compounds, especially structural analogues that interfere with the detection. In this work, a fluorescent covalent organic framework (named COF-TD), which can be used for the selective fluorescence detection and enrichment of myricetin from complex samples, was reported for the first time. The highly crystalline COF-TD with bright blue fluorescence was formed through a solution polymerization method by the condensation reaction between 4,4',4″-(1,3,5-triazine-2,4,6-triyl)trianiline and 2,5-dihydroxy-1,4-benzenedicarboxaldehyde. Due to spatial size selectivity, multisites hydrogen bonding, and π-π interaction, myricetin can quench the fluorescence of COF-TD with an inner filter effect (IFE) and static quenching mechanisms as well as can be enriched on COF-TD. Myricetin can observably eliminate the interference of other compounds and selectively quench the fluorescence of COF-TD with a limit of detection (LOD) of 0.30 µg·mL-1. The high adsorption ability of COF-TD (Q = 124.6 mg·g-1) to myricetin was also obtained. Finally, a sensing platform based on COF-TD for myricetin was successfully developed and applied for the detection of myricetin from vine teas. In addition, COF-TD also showed good water sensing ability and could be used effectively to detect water content in organic solvent (1-18% water in acetone, 0.5-5% water in acetonitrile, 1-4.5% water in ethyl acetate, v/v). To the best of our knowledge, this is the first report where COF-TD was used to detect water in a relatively wide concentration range. In all, this work provided dual-functional fluorescent COFs with the properties of an adsorbent, opening up new methodologies for the simple, selective, and enrichment detection method for myricetin.

Studies on the Mechanism of Alloimperatorin on the Proliferation and Apoptosis of HeLa Cells.

Alloimperatorin is a compound extracted from the traditional Chinese medicine (Angelica dahurica), which has exhibited anticancer activity. However, its precise molecular mechanism of anticancer remains unclear. Alloimperatorin-induced apoptosis of cervical cancer cells and its molecular mechanism were investigated in the present study. Cholecystokinin octapeptide (CCK-8) was employed to evaluate the cytotoxicity of alloimperatorin on HeLa, SiHa, and MS-751 cells. Flow cytometry was used to assess apoptosis induced by alloimperatorin. The mechanism of apoptosis was verified by mitochondrial membrane potential, Western blotting, and fluorescent PCR. The results of the study showed that alloimperatorin reduced the activity of HeLa cells. The calculated IC50 at 48 hours was 116.9 µM. Compared with the control group, alloimperatorin increased the apoptotic rate of HeLa cells and reduced the mitochondrial membrane potential of HeLa cells. The Western blot results showed that alloimperatorin promotes the expression of caspase3, 8, 9 and that Bax apoptotic proteins reduce PARP expression, procaspase3, 8, 9, and BCL-2 proteins and reduces the cyt-c in the mitochondria expression. The results demonstrated that alloimperatorin can induce HeLa cell apoptosis through mitochondria and extrinsic apoptotic pathways.

Scutellariabarbata D. Don extraction selectively targets stemness-prone NSCLC cells by attenuating SOX2/SMO/GLI1 network loop.

ETHNOPHARMACOLOGICAL RELEVANCE: Scutellariabarbata D. Don extraction (SBE), a traditional Chinese medicine, has been proved effective against various malignant disorders in clinics with tolerable side-effects when administered alone or in combination with conventional chemotherapeutic regimens. AIM OF THIS STUDY: Multi-drug resistance of cancer is attributed to existence of cancer stemness-prone cells that harbor aberrantly high activation of Sonic Hedgehog (SHH) cascade. Our previous study has demonstrated that SBE sensitized non-small cell lung cancer (NSCLC) cells to Cisplatin (DDP) treatment by downregulating SHH pathway. Yet, whether SBE could prohibit proliferation of cancer stemness-prone cells and its underlying molecular mechanisms remain to be investigated. In this article, we further investigated intervention of SBE on NSCLC cell stemness-associated phenotypes and its potential mode of action. MATERIALS AND METHODS: CCK-8 and clonal formation detection were used to measure the anti-proliferative potency of SBE against NSCLC and normal epithelial cells. Sphere formation assay and RQ-PCR were used to detect proliferation of cancer stemness cells and associated marker expression upon SBE incubation. Mechanistically, DARTS-WB and SPR were used to unveil binding target of SBE. Immunodeficient mice were implanted with patient derived tumor bulk for in vivo validation of anti-cancer effect of SBE. RESULTS: SBE selectively attenuated proliferation and stemness-like phenotypes of NSCLC cells rather than bronchial normal epithelial cells. Drug-protein interaction analysis revealed that SBE could directly bind with stem cell-specific transcription factor sex determining region Y-box 2 (SOX2) and interfere with the SOX2/SMO/GLI1 positive loop. In vivo assay using patient-derived xenografts (PDXs) model further proved that SBE diminished tumor growth and SOX2 expression in vivo. CONCLUSION: Our data indicate that SBE represses stemness-related features of NSCLC cells via targeting SOX2 and may serve as an alternative therapeutic option for clinic treatment.

Three-phase hollow fiber liquid-phase microextraction combined with HPLC for determination of three trace acidic plant growth regulators in Anoectochilus roxburghii (Wall.) Lindl.

The analysis of plant growth regulators presents a challenge due to their trace quantities and complex matrices. A novel, simple, and effective analytical method for the determination of three trace acidic plant growth regulators in Anoectochilus roxburghii (Wall.) Lindl was developed to address this issue. Three-phase hollow fiber liquid-phase microextraction combined with high-performance liquid chromatography was applied for the enrichment, purification, and determination of three acidic plant growth regulators, namely, indole-3-acetic-acid, indole-3-butyric-acid, and (+)-abscisic acid. The factors affecting extraction performance, including extractant species, pH of donor and acceptor phases, salt addition dosage, extraction time, temperature, and stirring rate, were investigated and optimized. Under optimum conditions, the proposed method provided good linearity (R2 , 0.9994-0.9999), low limit of detection (0.038-0.12 ng/mL), and acceptable relative recoveries (56.7-117.6%). The enrichment factors were between 153 and 328. The developed method was successfully applied to the enrichment and determination of plant growth regulators in Anoectochilus roxburghii (Wall.) Lindl and exhibited increased purification capacity, higher sensitivity, and decreased organic solvent consumption compared with conventional sample preparation methods. This method may provide a testing platform for the monitoring of plant growth regulator residues, ensuring the safe and effective use of traditional Chinese medicine.

Two new ent-atisane diterpenes from Sapium sebiferum.

Two new ent-atisane diterpenes, sebiferumnins K and L (1 and 2), were isolated from the branches of Sapium sebiferum, together with four known compounds, ent-3-oxoatis-16α, 17-acetonide (3), ent-l6α, 17-dihydroxyatisan-3-one (4), ent-atisane-3ß, 16α, 17-triol (5), ent-3α-hydroxyatis-16α,17-acetonide (6). The structures of these two ent-atisane diterpenes were elucidated by extensive analyses of the spectroscopic data. [Formula: see text].

[Clinical trials of treatment of woman menopause insomnia due to disharmony between heart and kidney by body and auricular acupuncture].

OBJECTIVE: To observe the therapeutic effect of body acupuncture combined with auricular acupuncture in the treatment of woman menopause insomnia induced by disharmony between the heart and kidney. METHODS: A total of 111 outpatients with menopausal insomnia were randomly divided into treatment group (n＝55, body acupoints plus auricular points) and control group (n＝56, body acupoints). For control group, Shenmen (HT7), Sanyinjiao (SP6), Anmian, Zhaohai (KI6), Shenmai (BL62), Sishencong (EX-HN1), Xinshu (BL15) and Shenshu (BL21) were needled with filiform needles, and HT7, SP6, Anmian, BL15 and BL21 were also stimulated with electroacupuncture (EA) for 30 min, once every other day, 3 times a week for 3 weeks. For the treatment group, auricular points as Kidney, Shenmen, Sympathetic and Endocrine around the ear canal and the ear cavity were punctured with filiform needles in combination with the same intervention methods mentioned above in the control group. The quality and efficiency of sleep were assessed by using Pittsburgh Sleep Quality Index (PSQI) scale before and after the treatment. The symptom scores of traditional Chinese medicine (TCM) and the therapeutic effect were determined according to the "Guide Principles for Clinical Research of New Drugs of TCM" (formulated by Chinese Ministry of Health, 1997). RESULTS: After the treatment, the scores of sleep quantity and effective rate of PSQI scale, and those of insomnia and dreaminess, dysphoria with feverish sensation in chest, palms and soles, paplpitation and irritability, fatigue and forgetfulness of TCM were reduced significantly in both groups compared with their own pre-treatment (P<0.05). Of the 56 and 55 cases in the control and treatment groups, 18 (32.1%) and 30 (54.5%) experienced a marked improvement in their symptoms, 28 (50.0%) and 22 (40.0%) were effective, and 10 (17.9%) and 3 (5.5%) failed, with the effective rate being 82.1% and 94.5%, respectively. The therapeutic effect of stimulation of the body acupoints plus auricular acupoints was significantly superior to that of stimulation of simple body acupoints in lowering scores of PSQI scale, insomnia and dreaminess, dysphoria with feverish sensation in chest, palms and soles, dizziness and tinninus, paplpitation and irritability, and tital fever and night sweating of TCM, as well as the effective rate (P<0.05). CONCLUSION: Both body acupuncture and body acupuncture combined with auricular acupuncture are effective in the treatment of woman menopause insomnia due to disharmony between the heart and kidney, and the two combined treatment was evidently better.

Magnetic dispersive solid-phase micro-extraction combined with high-performance liquid chromatography for determining nucleotides in Anoectochilus roxburghii (Wall.) Lindl.

A novel, simple, and efficacious analytical method for determining of nucleotides in Anoectochilus roxburghii (Wall.) Lindl (A. roxburghii) was developed. Magnetic dispersive solid-phase micro-extraction (MDSPME) combined with high-performance liquid chromatography was applied for extraction and determination of three nucleotides, such as adenosine-5'-monophosphate (AMP), uridine-5'-monophosphate (UMP) and guanosine-5'-monophosphate (GMP) in A. roxburghii from different sources. The structure and morphology of magnetic nanoparticles, Fe3O4@GO, were illustrated by Fourier-transform infrared (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM), and Thermagravimetric analysis (TGA) techniques. The effects of different extraction conditions on extraction efficiency were investigated and optimized. The optimum extraction conditions were performed as follows: 40.0 mg Fe3O4@GO were dispersed in 30 mL adsorption solution (pH 3.50, 2 µg/mL), 50 mM NaOH was employed for elution with 12 min of ultra-sonication at 40 °C. Under the aforementioned extraction conditions, the Fe3O4@GO nano-adsorbent obtained an excellent adsorption property. The corresponding linearity range for all three analytes exhibited a good linearity (r2 ≥ 0.9982) and notable added recoveries ranging from 88.4% to 109.8%, whereas the limit of quantitation was between 0.8-8 ng/mL. The enrichment factors (EF) were between 174 and 255. The proposed method showed the advantages of full purification, high EF, simplicity, and good recovery. The method was also successfully applied to nucleotides extraction and determination in A. roxburghii, showing superior reproducibility and high sensitivity. Based on this, the method could be expected to provide a novel experimental means and developmental direction for improving pretreatment and purification of nucleotides, reducing matrix effects as much as possible, in traditional Chinese medicinal materials or biological specimens.

The Alisma and Rhizoma decoction abates nonalcoholic steatohepatitis-associated liver injuries in mice by modulating oxidative stress and autophagy.

BACKGROUND: To investigate the effects of the Alisma and Rhizoma decoction on nonalcoholic steatohepatitis (NASH) and to further shed light on the underlying mechanisms of the actions of the Alisma and Rhizoma decoction. METHODS: Plasma alanine aminotransferase (ALT) content was determined and liver inflammation and fibrosis were evaluated. Intrahepatocellular malondialdehyde and superoxide dismutase contents were determined using commercially available kits Furthermore, α-SMA expression in liver tissues was examined by immunohistochemistry and LC3-II was detected by immunoblotting assays. RESULTS: Mice receiving the Alisma and Rhizoma decoction by gastric lavage had significantly lower plasma ALT content and markedly higher hepatic superoxide dismutase activity than mice receiving the methionine-choline deficient (MCD) diet. Furthermore, the decoction aborted MCD-induced increase in liver malondialdehyde content. Immunohistochemistry showed that the decoction suppressed hepatic α-SMA expression. Our transmission electronic microscopy revealed that the decoction markedly reduced the number of autophagosomes and immunoblotting assays showed that the decoction caused a dose-dependent decrease in LC3-II in hepatic tissues. CONCLUSION: The Alisma and Rhizoma decoction lessens NASH-associated liver injuries by modulating oxidative stress and autophagy in hepatocytes of mice fed with MCD.

[A new phenolic compound from endophytic fungus Aspergillus fumigatus of Euphorbia royleana].

This research was carried out to study the secondary metabolites of endophytic fungus Aspergillosis fumigatus from Euphorbia royleana. The endophytic fungus A. fumigatus was fermented by solid fermentation,and purified by various chromatographic methods after extraction. The structures of the compounds were identified by1 H-NMR,13 C-NMR and HSQC,HMBC spectra and physicchemical properties. Three compounds were isolated and their structures were identified as 3-( 3,4-dihydroxybenzoyl)-5-( 3,4-dihydroxyphenyl)-6-methyl-5,6-dihydro-2 H-pyran-2-one( 1),hydroxysydonic acid( 2) and 11-hydroxysydonic acid( 3). Compound 1 is a new compound.

Paeonin extracted from potatoes protects gastric epithelial cells from H2O2-induced oxidative damage in vitro by PI3K/Akt-mediated Nrf2 signaling pathway.

In this study, it is aimed to investigate the antioxidant mechanism of new extracts from potatoes. Four pigments, namely, Petunin, Paeonin, Malvidin and Pelargonidin, were extracted from potatoes by high performance liquid chromatography (HPLC). Our results showed that the cellular morphology and cell viability were significantly altered in gastric mucosal epithelial cells (GES-1) treated with different hydrogen peroxide (H2O2) concentrations over time (P < 0.05). Paeonin presented the strongest anti-oxidative effects on H2O2-treated cells, in both a dose- and time-dependent manner, determined by ARE-luciferase activity and HO-1 mRNA expression. After pre-treatment with Paeonin in H2O2-exposed cells, Keap1, Nrf2, HO-1 and NQO1 protein expressions were remarkably up-regulated. Furthermore, immunostaining of Nrf2 expression was obviously elevated in the H2O2 + Paeonin group over time. The GSH content in the H2O2 + Paeonin group was notably lower than that in the H2O2 + Paeonin + GSK690693 group. Paeonin promoted cell cycle with augmented Cyclin D1 and p27 protein expressions. Moreover, Paeonin suppressed apoptosis with increased Bcl2, total Caspase3 and total Caspase8 protein expressions and decreased Bax, p-Caspase3 and p-Caspase8 protein expression in H2O2-treated cells. These results suggested that Paeonin might exert an anti-oxidative role by activating Nrf2 signaling pathway with the changes of cell cycle and apoptosis.

Alisol A 24-acetate ameliorates nonalcoholic steatohepatitis by inhibiting oxidative stress and stimulating autophagy through the AMPK/mTOR pathway.

Alisol A 24-acetate (AA), a natural triterpenoid isolated from the traditional Chinese medicine Rhizoma Alismatis, has various therapeutic effects. We investigated the anti-nonalcoholic steatohepatitis (NASH) effect of AA and its underlying mechanisms in vitro and in vivo. C57BL/6 mice were fed a methionine and choline-deficient (MCD) diet for 4 weeks to induce NASH. The mice were simultaneously treated with a daily dose of AA (15, 30, and 60 mg kg-1, ig) for 4 weeks. On the last day, the animals were sacrificed and plasma and liver tissue were collected. Serum and liver tissue biochemical analyses and histological observation were performed. The human hepatic stellate cell line LX-2 was used to build NASH models by culturing with conditioned medium from WRL-68 liver cells after exposure to MCD medium in vitro. Liver oxidative stress and inflammatory indices and autophagy markers were examined. The results showed that AA suppressed reactive oxygen species (ROS) and inflammation in a NASH mouse model and inhibited the expression of inflammatory cytokines and ROS in LX-2 cells in MCD medium. Furthermore, we found AA stimulated autophagy in mice liver and LX-2, which could be the underlying mechanism of AA in NASH. To further investigate the role of autophagy in LX-2 cells, we found that AA regulated autophagy via the AMPK/mTOR/ULK1 pathway and dorsomorphin, a selective AMPK inhibitor, led to the suppression of AA-induced autophagy. Taken together, our results indicate that AA could be a possible therapy for NASH by inhibiting oxidative stress and stimulating autophagy.

Sodium selenite attenuates lung adenocarcinoma progression by repressing SOX2-mediated stemness.

PURPOSE: Sodium selenite (SS) has been widely reported to induce apoptosis in various cancer cell types. However, the underlying molecular mechanisms governing SS-mediated repression of lung cancer stem cells remain largely undefined. METHODS: In vitro assays of cell proliferation, clonal formation, apoptosis, migration and cancer stemness cell sphere formation were performed to examine the inhibitory effects of SS on lung adenocarcinoma (LAD) cells with or without the overexpression of SRY-related high-mobility-group box 2 (SOX2). RESULTS: SS significantly inhibited cell growth and induced apoptosis in LAD cells in a dose-dependent manner with marginal effects on normal epithelial cell HBEC. SS dramatically repressed expression of SOX2 and its upstream regulator GLI1 and strongly decreased stemness sphere formation in LAD cells at 10 µM. Forced expression of SOX2 significantly buffered anti-cancer effects of SS. CONCLUSIONS: Our results demonstrate that SS attenuates lung adenocarcinoma progression by repressing SOX2 and its upstream regulator GLI1, which suggests that SS may be a potential therapeutic drug candidate for lung cancer patients.

The MiR-135b-BMAL1-YY1 loop disturbs pancreatic clockwork to promote tumourigenesis and chemoresistance.

Circadian disruption has been implicated in tumour development, but the underlying mechanism remains unclear. Here, we show that the molecular clockwork within malignant human pancreatic epithelium is disrupted and that this disruption is mediated by miR-135b-induced BMAL1 repression. miR-135b directly targets the BMAL1 3'-UTR and thereby disturbs the pancreatic oscillator, and the downregulation of miR-135b is essential for the realignment of the cellular clock. Asynchrony between miR-135b and BMAL1 expression impairs the local circadian gating control of tumour suppression and significantly promotes tumourigenesis and resistance to gemcitabine in pancreatic cancer (PC) cells, as demonstrated by bioinformatics analyses of public PC data sets and in vitro and in vivo functional studies. Moreover, we found that YY1 transcriptionally activated miR-135b and formed a 'miR-135b-BMAL1-YY1' loop, which holds significant predictive and prognostic value for patients with PC. Thus, our work has identified a novel signalling loop that mediates pancreatic clock disruption as an important mechanism of PC progression and chemoresistance.

Disruption of SHH signaling cascade by SBE attenuates lung cancer progression and sensitizes DDP treatment.

Deregulated Sonic Hedgehog (SHH) pathway facilitates the initiation, progression, and metastasis of Non-small cell lung cancer (NSCLC), confers drug resistance and renders a therapeutic interference option to lung cancer patients with poor prognosis. In this study, we screened and evaluated the specificity of a Chinese herb Scutellariabarbata D. Don extraction (SBE) in repressing SHH signaling pathway to block NSCLC progression. Our study confirmed that aberrant activation of the SHH signal pathway conferred more proliferative and invasive phenotypes to human lung cancer cells. This study revealed that SBE specifically repressed SHH signaling pathway to interfere the SHH-mediated NSCLC progression and metastasis via arresting cell cycle progression. We also found that SBE significantly sensitized lung cancer cells to chemotherapeutic agent DDP via repressing SHH components in vitro and in vivo. Mechanistic investigations indicated that SBE transcriptionally and specifically downregulated SMO and consequently attenuated the activities of GLI1 and its downstream targets in SHH signaling pathway, which interacted with cell cycle checkpoint enzymes to arrest cell cycle progression and lead to cellular growth inhibition and migration blockade. Collectively, our results suggest SBE as a novel drug candidate for NSCLC which specifically and sensitively targets SHH signaling pathway.