Homoharringtonine inhibits the progression of hepatocellular carcinoma by suppressing the PI3K/AKT/GSK3ß/Slug signaling pathway.

Homoharringtonine (HHT), was first isolated from the bark of Cephalotaxus harringtonia (Knight ex J. Forbes) K. Koch and Cephalotaxus fortunei Hook trees. The bark extract is used to treat leukemia and in recent years has also been used in traditional Chinese medicine (TCM) to treat solid tumors. However, the inhibitory mechanism of HHT in the progression of hepatocellular carcinoma (HCC) is rarely studied. We aimed to evaluate the antitumor efficacy of HHT on HCC in vitro and in vivo and elucidate the underlying molecular mechanism(s). HCC cell lines, including HCCLM3, HepG2, and Huh7, were used to evaluate the antitumor efficacy of HHT in vitro. Cytotoxicity and proliferative ability were evaluated by MTT and colony formation assays. Cell cycle progression and apoptosis in HHT-treated HCC cells were evaluated by flow cytometry. To determine the migration and invasion abilities of HCC cells, wound-healing and Transwell assays were used. Finally, western blot analysis was used to reveal the proteins involved. We also established a xenograft nude mouse model for in vivo assessments of the preclinical efficacy of HHT, mainly using hematoxylin and eosin staining, immunohistochemistry, ultrasound imaging (USI), and magnetic resonance imaging (MRI). HHT suppressed the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of HCC cells, and induced cell cycle arrest at the G2 phase and apoptosis. In the HCC xenograft model, HHT showed an obvious tumor-suppressive effect. Surprisingly, Slug expression was also decreased by HHT via the PI3K/AKT/GSK3ß signaling pathway at least partially suppressed the growth of HCC via the PI3K/AKT/GSK3ß/Slug signaling pathway.

Bioassay-guided isolation of a Mycobacterium tuberculosis bioflim inhibitor from Arisaema sinii Krause.

Mycobacterium tuberculosis biofilms harbour drug-tolerant bacteria. Identification of drugs that inhibit biofilm formation could enable the dramatic shortening of tuberculosis treatments using standard antibiotics. Arisaema sinii Krause is used to treat pulmonary and lymphatic tuberculosis by Dong People of China. Current study was aimed to purify the active components against M. tuberculosis biofilms from Arisaema sinii extract by using bioassay-guided isolation. (E)-2-(methyl (phenyl) amino) ethyl 2-(2-hydroxyundecanamido)-7, 11-dimethyl-3-oxotetradec-4-enoate, compound 1, was identified as the active component. It could inhibit mycobacterial biofilm formation, disperse the preformed biofilms, and disrupt the mature biofilms at concentration of 4, 8, and 32 µg/ml, respectively. At the dose of 32 µg/ml, it could potentiate the bactericidal activity of isoniazid against M. tuberculosis in mature biofilms. The results of this study indicate that compound 1 might be a novel lead compound against mycobacterial biofilm formation.

[Distribution and sequestration of straw carbon in surface and deep soil aggregates under different fertilization treatments]. / ä¸åŒæ–½è‚¥æ¡ä»¶ä¸‹ç§¸ç§†ç¢³åœ¨è¡¨å±‚å’Œæ·±å±‚åœŸå£¤å›¢èšä½“ä¸­çš„åˆ†é ä¸Žå›ºå­˜.

Long-term fertilization causes the differences in water, heat, nutrients and microbial activities between topsoil and deep soil, with consequences on the decomposition and turnover of straw carbon (C) in soils. At a long-term positioning experimental station in Shenyang Agricultural University, we mixed the topsoil (0-20 cm) and deep soil (40-60 cm) samples from different fertilization treatments with 13C-labeled straw for in-situ incubation. We analyzed the content of organic C and its δ13C value in soil aggregates, compared the difference in the distribution of straw C between topsoil and deep soil aggregates, and explored the effects of fertilization on the sequestration of straw C in soil aggregates. Compared with fertilization treatments (i.e., single chemical nitrogen fertilizer application and combination of organic manure with nitrogen fertilizer application), the treatment without fertilization increased the content of straw C of <0.053 mm aggregate in the topsoil by 106.7% and that of >0.25 mm aggregate in the deep soil by 34.2%. The contribution percentage of straw C to organic C of >0.053 mm aggregate in the deep soil was about two times of that in the topsoil. About 22.6% and 11.4% of straw C was distributed into the >0.25 mm and <0.25 mm aggregates of topsoil, and about 29.4% and 8.8% of straw C was distributed into the >0.25 mm and <0.25 mm aggregates of deep soil, respectively. In conclusion, straw addition promoted the regeneration and sequestration of carbon in deep soil macroaggregates and increased the carbon sequestration potential of deep soil.

Construction of a prognostic immune signature for lower grade glioma that can be recognized by MRI radiomics features to predict survival in LGG patients.

BACKGROUND: This study aimed to identify a series of prognostically relevant immune features by immunophenoscore. Immune features were explored using MRI radiomics features to prediction the overall survival (OS) of lower-grade glioma (LGG) patients and their response to immune checkpoints. METHOD: LGG data were retrieved from TCGA and categorized into training and internal validation datasets. Patients attending the First Affiliated Hospital of Harbin Medical University were included in an external validation cohort. An immunophenoscore-based signature was built to predict malignant potential and response to immune checkpoint inhibitors in LGG patients. In addition, a deep learning neural network prediction model was built for validation of the immunophenoscore-based signature. RESULTS: Immunophenotype-associated mRNA signatures (IMriskScore) for outcome prediction and ICB therapeutic effects in LGG patients were constructed. Deep learning of neural networks based on radiomics showed that MRI radiomic features determined IMriskScore. Enrichment analysis and ssGSEA correlation analysis were performed. Mutations in CIC significantly improved the prognosis of patients in the high IMriskScore group. Therefore, CIC is a potential therapeutic target for patients in the high IMriskScore group. Moreover, IMriskScore is an independent risk factor that can be used clinically to predict LGG patient outcomes. CONCLUSIONS: The IMriskScore model consisting of a sets of biomarkers, can independently predict the prognosis of LGG patients and provides a basis for the development of personalized immunotherapy strategies. In addition, IMriskScore features were predicted by MRI radiomics using a deep learning approach using neural networks. Therefore, they can be used for the prognosis of LGG patients.

[Effects of long-term fertilization on soil ammonia-oxidizing microorganisms]. / 长期施肥对土壤氨氧化微生物的影响.

Long-term fertilization can change the supply of soil carbon and nitrogen (N), with consequences on the abundance and community structure of soil microorganisms. Based on the long-term fertilization positioning experiment station of brown earth, we analyzed the dynamics of soil ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) under different fertilization treatments, including no fertilization (CK), low level of inorganic N fertilizer (N2), high level of inorganic N fertilizer (N4), and organic manure combined with inorganic N fertilizer (M2N2), aiming to provide a basis for microbiological mechanism of soil N transformation and improvement of soil fertility. The results showed that the ratio of AOA to AOB abundance was 2.28-61.95 under different fertilization treatments. Compared with that in CK, the AOA abundance was reduced by 1.6%-13.6% after long-term fertilization. The abundance of AOB in N4 treatment decreased first and then increased with soil depths, but with contrary results in other treatments. The Shannon diversity index (H), evenness index (J), and Simpson index (S) of AOB were higher than those of AOA. The AOB diversity was increased at 0-20 cm soil layer in M2N2 treatment, while that of AOA was decreased. Soil AOB clustered with soil depths, and neither AOA nor AOB community clustered with fertilization treatments. In summary, long-term fertilization altered the composition of AOA and AOB. AOA was sensitive to environment, whereas AOB was more abundant and stable.

Feature-Based Molecular Networking Analysis of the Metabolites Produced by In Vitro Solid-State Fermentation Reveals Pathways for the Bioconversion of Epigallocatechin Gallate.

Dark teas are prepared by a microbial fermentation process. Flavan-3-ol B-ring fission analogues (FBRFAs) are some of the key bioactive constituents that characterize dark teas. The precursors and the synthetic mechanism involved in the formation of FBRFAs are not known. Using a unique solid-state fermentation system with ß-cyclodextrin inclusion complexation as well as targeted chromatographic isolation, spectroscopic identification, and Feature-based Molecular Networking on the Global Natural Products Social Molecular Networking web platform, we reveal that dihydromyricetin and the FBRFAs, including teadenol A and fuzhuanin A, are derived from epigallocatechin gallate upon exposure to fungal strains isolated from Fuzhuan brick tea. In particular, the strains from subphylum Pezizomycotina were key drivers for these B-/C-ring oxidation transformations. These are the same transformations seen during the fermentation process of dark teas. These discoveries set the stage to enrich dark teas and other food products for these health-promoting constituents.

Oxidized epigallocatechin gallate inhibited lysozyme fibrillation more strongly than the native form.

Epigallocatechin gallate (EGCG), the most abundant flavanoid in green tea, is currently being evaluated in the clinic due to its benefits in the treatment of amyloid disorders. Its anti-amyloidogenic effect has been attributed to direct interaction of the intact molecule with misfolded polypeptides. In addition, antioxidant activity is also involved in the anti-amyloidogenic role. The detailed molecular mechanism is still unclear and requires further investigation. In the present study, the kinetics of EGCG oxidation and the anti-amyloidogenic effect of the resultant oxidation substances have been examined. The results indicate that EGCG degrades in a medium at pH 8.0 with a half-life less than 2h. By utilizing lysozyme as an in vitro model, the oxidized EGCG demonstrates a more potent anti-amyloidogenic capacity than the intact molecule, as shown by ThT and ANS fluorescence, TEM determination, and hemolytic assay. The oxidized EGCG also has a stronger disruptive effect on preformed fibrils than the native form. Ascorbic acid eliminates the disruptive role of native EGCG on the fibrils, suggesting that oxidation is a prerequisite in fibril disruption. The results of this work demonstrate that oxidized EGCG plays a more important role than the intact molecule in anti-amyloidogenic activity. These insights into the action of EGCG may provide a novel route to understand the anti-amyloidogenic activity of natural polyphenols.

Hyperthermia with different temperatures inhibits proliferation and promotes apoptosis through the EGFR/STAT3 pathway in C6 rat glioma cells.

Malignant gliomas are a group of aggressive neoplasms among human cancers. The curative effects of current treatments are finite for improving the prognosis of patients. Hyperthermia (HT) is an effective treatment for cancers; however, the effects of HT with different temperatures in treatment of MG and relevant mechanisms remain unclear. MTT assay and Annexin V­fluorescein isothiocyanate/propidium iodide staining were used for investigating the proliferation and apoptosis of C6 cells, respectively. Western blotting was applied to detect the expression of proteins. Ultrasonography was employed to evaluate the tumor formation rate, growth rate, angiogenesis rate and degree of hardness of tumors in vivo. The authors certified that HT with 42­46˚C x 1 h, 1 t could inhibit proliferation, promote apoptosis, reduce tumor formation rate, growth rate, angiogenesis rate, degree of hardness of tumors, ischemic tolerance and anoxic tolerance, and have synergy with temozolomide in C6 cells. Long­term HT (43˚C x 1 h, 1 t/5 d, 90 d) did not cut down the sensitivity of C6 cells to HT, and sustainably inhibited the proliferation of C6 cells. Furthermore, the authors proved HT produced these effects primarily through inhibition of the EGFR/STAT3/HIF­1A/VEGF­A pathway.

Protective Effects of Hydrogen Gas on Experimental Acute Pancreatitis.

Acute pancreatitis (AP) is an inflammatory disease mediated by damage to acinar cells and pancreatic inflammation. In patients with AP, subsequent systemic inflammatory responses and multiple organs dysfunction commonly occur. Interactions between cytokines and oxidative stress greatly contribute to the amplification of uncontrolled inflammatory responses. Molecular hydrogen (H2) is a potent free radical scavenger that not only ameliorates oxidative stress but also lowers cytokine levels. The aim of the present study was to investigate the protective effects of H2 gas on AP both in vitro and in vivo. For the in vitro assessment, AR42J cells were treated with cerulein and then incubated in H2-rich or normal medium for 24 h, and for the in vivo experiment, AP was induced through a retrograde infusion of 5% sodium taurocholate into the pancreatobiliary duct (0.1 mL/100 g body weight). Wistar rats were treated with inhaled air or 2% H2 gas and sacrificed 12 h following the induction of pancreatitis. Specimens were collected and processed to measure the amylase and lipase activity levels; the myeloperoxidase activity and production levels; the cytokine mRNA expression levels; the 8-hydroxydeoxyguanosine, malondialdehyde, and glutathione levels; and the cell survival rate. Histological examinations and immunohistochemical analyses were then conducted. The results revealed significant reductions in inflammation and oxidative stress both in vitro and in vivo. Furthermore, the beneficial effects of H2 gas were associated with reductions in AR42J cell and pancreatic tissue damage. In conclusion, our results suggest that H2 gas is capable of ameliorating damage to the pancreas and AR42J cells and that H2 exerts protective effects both in vitro and in vivo on subjects with AP. Thus, the results obtained indicate that this gas may represent a novel therapy agent in the management of AP.

[Effects of manure application on organic carbon in aggregates of black soil].

In this paper, black soil samples were separated into four sizes ( > 2,000, 2,000-250, 250-53, and <53 microm) of aggregates by wet sieving, and their free light fraction (free LF) and intra-aggregate particulate organic matter (iPOM) were isolated by density fractionation, aimed to evaluate the effects of manure application on the organic carbon in different sizes of aggregates in black soil. The results showed that compared with the control, applying manure improved soil aggregation significantly, and compensated the disturbance of tillage and slowed down the turnover of aggregates to some degree. After applying manure, the content of fine iPOM-C was significantly higher than that of coarse iPOM-C, suggesting that manure application was beneficial to the accumulation of fine iPOM-C, which is the main form of carbon sequestration in the aggregates of black soil.