Tumour metastasis is a major reason accounting for the poor prognosis of colorectal cancer (CRC), and the discovery of targets in the primary tumours that can predict the risk of CRC metastasis is ...now urgently needed. In this study, we identified autophagy-related protein 9B (ATG9B) as a key potential target gene for CRC metastasis. High expression of ATG9B in tumour significantly increased the risk of metastasis and poor prognosis of CRC. Mechanistically, we further find that ATG9B promoted CRC invasion mainly through autophagy-independent manner. MYH9 is the pivotal interacting protein for ATG9B functioning, which directly binds to cytoplasmic peptide segments aa368-411 of ATG9B by its head domain. Furthermore, the combination of ATG9B and MYH9 enhance the stability of each other by decreasing their binding to E3 ubiquitin ligase STUB1, therefore preventing them from ubiquitin-mediated degradation, which further amplified the effect of ATG9B and MYH9 in CRC cells. During CRC cell invasion, ATG9B is transported to the cell edge with the assistance of MYH9 and accelerates focal adhesion (FA) assembly through mediating the interaction of endocytosed integrin β1 and Talin-1, which facilitated to integrin β1 activation. Clinically, upregulated expression of ATG9B in human CRC tissue is always accompanied with highly elevated expression of MYH9 and associated with advanced CRC stage and poor prognosis. Taken together, this study highlighted the important role of ATG9B in CRC metastasis by promoting focal adhesion assembly, and ATG9B together with MYH9 can provide a pair of potential therapeutic targets for preventing CRC progression.
Rationale
Ginger pulp is the dried rhizome scraped off the skin which originates from Zingiber officinale Rosc., a Zingiberaceae plant. Ginger peel is the dried rhizome skin of Zingiber officinale ...Rosc. (Zingiberaceae). The present work aims to investigate the different chemical constituents that are related to the medicinal properties of the ginger pulp and ginger peel.
Methods
A rapid ultra‐high‐performance liquid chromatography/electrospray ionization quadrupole time‐of‐flight tandem mass spectrometry (UHPLC/ESI‐QTOF/MS) method was developed for qualitative analysis of the constituents in different polarity extracted fractions of the pulp and peel of ginger rhizomes.
Results
A total of 83 compounds were identified from the pulp and peel of ginger rhizomes, including 36 diarylheptanoids, 25 gingerols and 22 other compounds. Nine of these were new compounds. In total, 46, 27, 65 and 51 compounds were identified from the crude extract, petroleum ether, ethyl acetate, and n‐butanol fractions of the ginger pulp, respectively, and 60, 30, 70 and 62 compounds were identified from the crude extract, petroleum ether, ethyl acetate, n‐butanol fractions of the ginger peel, respectively. Each identified compound is marked on the corresponding chromatogram.
Conclusions
The integrated method is sensitive and reliable for searching the different chemical constituents from different polarity extracted fractions of the ginger pulp and ginger peel. This work may provide a significant contribution to research into the medicinal properties of the ginger pulp and ginger peel.
The fall armyworm (FAW), Spodoptera frugiperda Smith (Lepidoptera: Noctuidae) is an important agricultural pest that invaded China in the middle of December 2018. As a polyphagous pest, FAW is ...identified as a serious threat to agricultural production and food security in China. Pepper (Capsicum annuum L.), tomato (Solanum lycopersicum Mill.) and eggplant (Solanum melongena L.) are three of dominant solanaceous vegetables of this country. To our knowledge, the effects of these plants on the performances of FAW have not been well studied. In this study we assessed the fitness of this pest to these three plants. Results showed that FAW can complete its life cycle when fed with tomato and pepper, but not on eggplant. The population parameters of FAW fed with maize (Zea mays L.) and the three solanaceous vegetables were compared using the age-stage, two-sex life table method. Developmental duration was significantly different in the larval stage, but not in the pupae stage. FAW fed with pepper had the longest pre-adult period (41.73 d) and the lightest pupal weight (0.1134 g); the survival rate was lower than FAW fed with tomato. Significant differences were observed in the mean fecundity of female, with the highest (943.95 eggs) laid by FAW fed with tomato. FAW had the shortest mean generation time (T), the highest intrinsic rate of increase (r) and finite rate of increase (λ) on maize, and the highest net reproductive rate (R0) on tomato. Overall, FAW fitness on the three solanaceous vegetables was: tomato>pepper>eggplant. This study provides the foundation for further assessment of FAW risk to solanaceous vegetables and for establishing corresponding control strategies in China.
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A novel magnetic nanomaterial was synthesized by grafting nitrilotriacetic acid to magnetic graphene oxide (NDMGO), which was applied as an adsorbent for removing tetracycline (TC) ...from aqueous solutions. The nanomaterial was characterized using TG-DTA, SEM, TEM, XRD, VSM, XPS, Raman, BET surface area and zeta potential measurements. Several experimental conditions (solution pH, adsorption time, temperature, ionic strength and foreign ions) affecting the adsorption process were investigated. The results showed that the TC adsorption capacity could be affected by solution pH. The adsorption capacity of TC increased rapidly in the initial 20min and finally reached equilibrium was about 600min. The pseudo-second-order kinetics provided the better correlation for the experiment data. Various thermodynamic parameters indicated that the adsorption was a spontaneous and endothermic process. The presence of NaCl and background electrolytes in the solution had a slight influence on TC adsorption. Hydrogen bonds, amidation reaction, π-π and cation-π interaction between NDMGO and TC could be used to explain the adsorption mechanism. The regeneration experiment demonstrated that this nanomaterial possessed an excellent regeneration performance. Based on the experimental results and comparative analysis with other adsorbents, the NDMGO was a high-efficiency and reusable adsorbent for TC pollution control.
Tumor budding is included in the routine diagnosis of colorectal cancer (CRC) and is considered a tumor prognostic factor independent of TNM staging. This study aimed to identify the ...fibroblast-mediated effect of tumor bud-derived C-C chemokine ligand 5 (CCL5) on the tumor microenvironment (TME).
Recruitment assays and a human cytokine array were used to detect the main cytokines that CRC tumor buds secrete to recruit fibroblasts. siRNA transfection and inhibitor treatment were used to investigate the role of fibroblast CCL5 receptors in fibroblast recruitment. Subsequently, transcriptome sequencing was performed to explore the molecular changes occurring in fibroblasts upon stimulation with CCL5. Finally, clinical specimens and orthotopic xenograft mouse models were studied to explore the contribution of CCL5 to angiogenesis and collagen synthesis.
Hematoxylin-eosin staining and immunochemistry revealed a higher number of fibroblasts at the invasive front of CRC tissue showing tumor budding than at sites without tumor budding. In vitro experiments demonstrated that CCL5 derived from tumor buds could recruit fibroblasts by acting on the CCR5 receptors on fibroblasts. Tumor bud-derived CCL5 could also positively regulate solute carrier family 25 member 24 (SLC25A24) expression in fibroblasts, potentially activating pAkt-pmTOR signaling. Moreover, CCL5 could increase the number of α-SMA
CD90
FAP
fibroblasts and thus promote tumor angiogenesis by enhancing VEGFA expression and making fibroblasts transdifferentiate into vascular endothelial cells. Finally, the results also showed that CCL5 could promote collagen synthesis through fibroblasts, thus contributing to tumor progression.
At the invasive front of CRC, tumor bud-derived CCL5 can recruit fibroblasts via CCR5-SLC25A24 signaling, further promoting angiogenesis and collagen synthesis via recruited fibroblasts, and eventually create a tumor-promoting microenvironment. Therefore, CCL5 may serve as a potential diagnostic marker and therapeutic target for tumor budding in CRC.
Pancreatic ductal adenocarcinoma (PDAC), cancer with a high mortality rate and the highest rate of KRAS mutation, reportedly internalizes proteins via macropinocytosis to adapt to low amino acid ...levels in the tumor microenvironment. Here, we aimed to identify a key regulator of macropinocytosis for the survival of tumor cells in a low amino acid environment in PDAC. FYVE, RhoGEF, and PH domain-containing protein 6 (FGD6) were identified as key regulators of macropinocytosis. FGD6 promoted PDAC cell proliferation, macropinocytosis, and tumor growth both in vitro and in vivo. The macropinocytosis level was decreased with FGD6 knockdown in PDAC cell lines. Moreover, FGD6 promoted macropinocytosis by participating in the trans-Golgi network and enhancing the membrane localization of growth factor receptors, especially the TGF-beta receptor. TGF-beta enhanced macropinocytosis in PDAC cells. Additionally, YAP nuclear translocation induced by a low amino acid tumor environment initiated FGD6 expression by coactivation with YY1. Clinical data analysis based on TCGA and GEO datasets showed that FGD6 expression was upregulated in PDAC tissue, and high FGD6 expression was correlated with poor prognosis in patients with PDAC. In tumor tissue from Kras
/Trp53
/Pdx1-Cre (KPC) mice, FGD6 expression escalated during PDAC development. Our results uncover a previously unappreciated mechanism of macropinocytosis in PDAC. Strategies to target FGD6 and growth factors membrane localization might be developed for the treatment of PDAC.
Our previous studies have shown that the 3′ end of metastasis associated lung adenocarcinoma transcript 1 (MALAT1) is involved in colorectal cancer (CRC) cell proliferation and migration/invasion in ...vitro. The role and mechanism of MALAT1 in CRC metastasis in vivo, however, remain largely unknown. In the present study, we found that MALAT1 was up-regulated in human primary CRC tissues with lymph node metastasis. Overexpression of MALAT1 via RNA activation promoted CRC cell proliferation, invasion and migration in vitro, and stimulated tumor growth and metastasis in mice in vivo. Conversely, knockdown of MALAT1 inhibited CRC tumor growth and metastasis. MALAT1 regulated at least 243 genes in CRC cells in a genome-wide expression profiling. Among these genes, PRKA kinase anchor protein 9 (AKAP-9) was significantly up-regulated at both mRNA and protein levels. AKAP-9 was highly expressed in CRC cells with metastatic potential and human primary CRC tissues with lymph node metastasis, but not in normal cells or tissues. Importantly, knockdown of AKAP-9 blocked MALAT1-mediated CRC cell proliferation, migration and invasion. These data indicate that MALAT1 may promote CRC tumor development via its target protein AKAP-9.
•MALAT1 is up-regulated in human primary CRC tissues with lymph node metastasis.•MALAT1 can be induced via RNA activation.•MALAT1 plays a critical role in CRC tumor growth and metastasis in mice.•AKAP-9 is correlated with MALAT1 in CRC tissues.•AKAP-9 mediates MALAT1 function in CRC proliferation/migration/invasion.
Objective
Osteonecrosis of the femoral head (ONFH) is a common orthopedic disease with a high disability rate. The clinical effect of BuShenHuoXue decoction (BSHX) for ONFH is satisfactory. We aimed ...to elucidate the potential angiogenic mechanisms of BSHX in a rat femoral osteonecrosis model and bone marrow mesenchymal stem cells (BMSCs).
Methods
With in vivo experiments, we established the steroid‐induced osteonecrosis of the femoral head (SONFH) model using Sprague–Dawley (SD) rats (8‐week‐old). The rats were randomly divided into five group of 12 rats each and given the corresponding interventions: control, model (gavaged with 0.9% saline), BSHX low‐, medium‐ and high‐dose groups (0.132 3, 0.264 6, and 0.529 2 g/mL BSHX solution by gavage). After 12 weeks, haematoxylin and eosin (H&E) staining was preformed to evaluate rat osteonecrosis. the expression of angiogenic factors (CD31, VEGFA, KDR, VWF) in rat femoral head was detected by immunohistochemistry, qPCR and western blotting. In cell experiment, BMSCs were isolated and cultured in the femoral bone marrow cavity of 4‐week‐old SD rats. BMSCs were randomly divided into eight groups and intervened with different doses of BSHX‐containing serum and glucocorticoids: control group (CG); BSHX low‐, medium‐, and high‐dose groups (CG + 0.661 5, 1.323, and 2.646 g/kg BSHX gavage rat serum); dexamethasone (Dex) group; and Dex + BSHX low‐, medium‐, and high‐dose groups (Dex + 0.661 5, 1.323, and 2.646 g/kg BSHX gavaged rat serum), the effects of BSHX‐containing serum on the angiogenic capacity of BMSCs were examined by qPCR and Western blotting. A co‐culture system of rat aortic endothelial cells (RAOECs) and BMSCs was then established. Migration and angiogenesis of RAOECs were observed using angiogenesis and transwell assay. Identification of potential targets of BSHX against ONFH was obtained using network pharmacology.
Results
BSHX upregulated the expression of CD31, VEGFA, KDR, and VWF in rat femoral head samples and BMSCs (p < 0.05, vs. control group or model group). Different concentrations of BSHX‐containing serum significantly ameliorated the inhibition of CD31, VEGFA, KDR and VWF expression by high concentrations of Dex. BSHX‐containing serum‐induced BMSCs promoted the migration and angiogenesis of RAOECs, reversed to some extent the adverse effect of Dex on microangiogenesis in RAOECs, and increased the number of microangiogenic vessels. Furthermore, we identified VEGFA, COL1A1, COL3A1, and SPP1 as important targets of BSHX against ONFH.
Conclusion
BSHX upregulated the expression of angiogenic factors in the femoral head tissue of ONFH model rats and promoted the angiogenic capacity of rat RAOECs and BMSCs. This study provides an important basis for the use of BSHX for ONFH prevention and treatment.
BSHX ameliorated glucocorticoid‐induced decrease in angiogenesis in the rat femoral head. BSHX‐containing serum promoted the migratory proliferative activity of rat aortic endothelial cells (RAOECs) and the angiogenic differentiation ability of bone marrow mesenchymal stem cells (BMSCs). VEGFA, COL1A1, COL3A1, and SPP1 were critical targets of BSHX in the prevention and treatment of osteonecrosis of the femoral head (ONFH).
The screening performance of urine flow cytometry parameters (e.g., white blood cell and bacteria) for urinary tract infection (UTI) has been widely recognized. The majority of previous studies, ...however, investigated the screening performance of Sysmex UF-1000i urine flow cytometer. This study aimed to investigate the screening performance of Sysmex UF-5000 analyzer, a third-generation urinary flow cytometer, for UTI and its novel parameter named Gram flag for discriminating gram-positive and negative pathogens. Urine specimens sent to the clinical microbiology laboratory of our hospital for bacterial culture between September 13, 2021, and November 15, 2021, were prospectively and consecutively collected. The Sysmex UF-5000 analyzer was used to determine urine white blood cell (WBC) and bacteria simultaneously. A chemical strip was used to assess urine nitrate. UTI was defined as positive urine bacterial culture > 104 CFU /ml. The receiver operating characteristics (ROC) curve, nomogram, decision tree, and decision curve were used to determine the screening performance of urine WBC, nitrate, and bacterial. A total of 246 UTIs and 425 non-UTIs were enrolled. The areas under the ROC curve (AUCs) for WBC and bacterial were 0.74 and 0.86, respectively. The decision curve showed that urine bacteria had a higher benefit than WBC. The nomogram indicated that urine bacterial had the largest effect on the probability of UTI. The sensitivity and specificity of the decision tree were 0.69 and 0.95, respectively. The flag of Gram-negative had a positive predictive value (PPV) of 0.93 in patients with urine bacteria > 1367 /μl. Therefore, we conclude that urine bacteria determined by the Sysmex UF-5000 had higher screening performance and greater benefit than WBC. The decision tree can be used to improve the screening performance of routine urinary parameters. The flag of Gram-negative is a reliable indicator to confirm gram-negative bacteria infection in UTI patients.
In our research, we explored a natural substance called Oxymatrine, found in a traditional Chinese medicinal plant, to fight against a common bird flu virus known as H9N2. This virus not only affects ...birds but can also pose a threat to human health. We focused on how this natural compound can help in stopping the virus from spreading in cells that line the lungs of birds and potentially humans. Our findings show that Oxymatrine can both directly block the virus and boost the body's immune response against it. This dual-action mechanism is particularly interesting because it indicates that Oxymatrine might be a useful tool in developing new ways to prevent and treat this type of bird flu. Understanding how Oxymatrine works against the H9N2 virus could lead to safer and more natural ways to combat viral infections in animals and humans, contributing to the health and well-being of society. The H9N2 Avian Influenza Virus (AIV) is a persistent health threat because of its rapid mutation rate and the limited efficacy of vaccines, underscoring the urgent need for innovative therapies. This study investigated the H9N2 AIV antiviral properties of Oxymatrine (OMT), a compound derived from traditional Chinese medicine, particularly focusing on its interaction with pulmonary microvascular endothelial cells (PMVECs). Employing an array of in vitro assays, including 50% tissue culture infectious dose, Cell Counting Kit-8, reverse transcription-quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blot, we systematically elucidated the multifaceted effects of OMT. OMT dose-dependently inhibited critical antiviral proteins (PKR and Mx1) and modulated the expression of type I interferons and key cytokines (IFN-α, IFN-β, IL-6, and TNF-α), thereby affecting TLR3 signaling and its downstream elements (NF-κB and IRF-3). OMT's antiviral efficacy extended beyond TLR3-mediated responses, suggesting its potential as a versatile antiviral agent. This study not only contributes to the growing body of research on the use of natural compounds as antiviral agents but also underscores the importance of further investigating the broader application of OMT for combating viral infections.
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