The truncated AXIN1 isoform promotes hepatocellular carcinoma metastasis through SRSF9-mediated exon 9 skipping.

Axis inhibitor protein 1 (AXIN1) is a protein recognized for inhibiting tumor growth and is commonly involved in cancer development. In this study, we explored the potential molecular mechanisms that connect alternative splicing of AXIN1 to the metastasis of hepatocellular carcinoma (HCC). Transcriptome sequencing, RT‒PCR, qPCR and Western blotting were utilized to determine the expression levels of AXIN1 in human HCC tissues and HCC cells. The effects of the AXIN1 exon 9 alternative splice isoform and SRSF9 on the migration and invasion of HCC cells were assessed through wound healing and Transwell assays, respectively. The interaction between SRSF9 and AXIN1 was investigated using UV crosslink RNA immunoprecipitation, RNA pulldown, and RNA immunoprecipitation assays. Furthermore, the involvement of the AXIN1 isoform and SRSF9 in HCC metastasis was validated in a nude mouse model. AXIN1-L (exon 9 including) expression was downregulated, while AXIN1-S (exon 9 skipping) was upregulated in HCC. SRSF9 promotes the production of AXIN1-S by interacting with the sequence of exons 8 and 10 of AXIN1. AXIN1-S significantly promoted HCC cells migration and invasion by activating the Wnt pathway, while the opposite effects were observed for AXIN1-L. In vivo experiments demonstrated that AXIN1-L inhibited HCC metastasis, whereas SRSF9 promoted HCC metastasis in part by regulating the level of AXIN1-S. AXIN1, a tumor suppressor protein that targets the AXIN1/Wnt/ß-catenin signaling axis, may be a promising prognostic factor and a valuable therapeutic target for HCC.

Sarsasapogenin-AA13 inhibits LPS-induced inflammatory responses in macrophage cells in vitro and relieves dimethylbenzene-induced ear edema in mice.

Sarsasapogenin-AA13 (AA13) is a novel synthetic derivative of sarsasapogenin extracted from the Chinese herb Rhizoma Anemarrhenae. In this study we investigated the effects of AA13 on lipopolysaccharide (LPS)-induced production of inflammatory factors in macrophage cells and the anti-inflammatory activity of AA13 in an inflammatory model of dimethylbenzene-induced ear edema. Macrophage cells (RAW264.7 cells and mouse peritoneal macrophages) were exposed to LPS (1 µg/mL); pretreatment with AA13 (5-20 µmol/L) dose-dependently inhibited LPS-induced production of NO, TNF-α and PGE2, and LPS-stimulated expression levels of COX-2 and iNOS. Furthermore, pretreatment with AA13 dose-dependently suppressed LPS-stimulated phosphorylation of p38 and JNK, but had no effect on ERK in RAW264.7 cells. Moreover, pretreatment with AA13 inhibited LPS-induced activation of the nuclear factor (NF)-κB in RAW264.7 cells. The in vivo anti-inflammatory activity of AA13 was demonstrated in a mouse inflammatory model: pre-treatment with either AA13 (20 mg·kg-1·d-1, ig) or a positive control antifani (10 mg·kg-1·d-1, ig) for 3 d significantly relieved dimethylbenzene-induced ear edema. Our results demonstrate that AA13 effectively inhibit LPS-induced inflammatory responses in macrophage cells in vitro and relieve dimethylbenzene-induced ear edema in vivo.

[Comparison and application of calculation methods for water consumption of tree transpiration]. / æž—æœ¨è’¸è ¾è€—æ°´é‡æµ‹ç®—æ–¹æ³•çš„æ¯”è¾ƒä¸Žåº”ç”¨.

In this study, the concept, research value, and research advances of tree transpiration were concisely narrated. From the perspective of measurement and estimation, we summarized the main calculation methods for water consumption of tree transpiration. By comparing the advantages, disadvantages, applicability, limitations, application status, and applicable scale of different methods, we showed that the measurement methods could be applied to the measurement of water consumption at multiple spatial scales, and that the estimation methods were often applied to the measurement of evapotranspiration at large spatial scales. As the measurement method was the basis of the estimation method, it is necessary to carry out reasonable quality control and evaluation of the measured results of measurement methods, in order to provide data basis for the estimation results of the correction estimation method. Whether it was at a particular large spatial scale or at different spatial scales, the combination of measurement methods and estimation methods could improve the accuracy of evapotranspiration measurement. Improving the accuracy of calculating the dispersion of trees at a large spatial scale in complex underlying surface and harsh climate environments would become a research hotspot but may also be a difficulty in the future. With the continuous progress of science and technology, both measurement devices and methods would be improved, and new methods for accurately measuring the water consumption of forest transpiration would be available.

Kinetic stability studies of HBV vaccine in a microneedle patch.

This study systematically demonstrated the antigenicity kinetics of HBV vaccine microneedles (MNs) during the fabrication, application and storage. To improve the stability of HBsAg in a microneedle patch, several selected saccharides were added to the MN formulations as stabilizers. According to the experimental data, no significant decrease of the bio-activity of HBsAg antigen was found during the microneedle fabrication process. And then immune effects of HBsAg added with different sugars were tested. Chitosan and trehalose loaded HBsAg MNs enhanced the antibody levels to approximately 1.5-fold and 2-fold of the plain HBsAg MNs respectively while sucrose and glucose were not obviously beneficial. During the short-term storage under 60 °C, the antigenicity of HBsAg MNs encapsulated with glucose and chitosan declined sharply in 24 h and hardly left after 7 days. As for the groups of HBsAg MNs added with sucrose and trehalose, approximately 90% of HBsAg initial antigenicity maintained, which could be attributed to the protective function of non-reductive disaccharides. As for the long-term storage experiments, the pharmacological activity of HBsAg antigen protected by sucrose and trehalose slightly reduced in 3 months except for the samples under 60 °C. In extreme condition, trehalose performed even better protection function than sucrose, of which the antigenicity of HBsAg in MNs left approximately 81% and 63% of its initial, respectively. These results confirmed that trehalose loaded HBsAg MNs enabled stable encapsulation and storage of HBsAg antigen and realized reasonable enhancement of immune effect in a relatively painless, safe, and convenient manner.

Icariin possesses chondroprotective efficacy in a rat model of dexamethasone-induced cartilage injury through the activation of miR-206 targeting of cathepsin K.

The present study aimed to investigate the articular cartilage and chondrocytes of dexamethasone (DXM)-induced cartilage injuries in rats in response to treatment with icariin, as well as the implicated molecular mechanism. Effects of icariin on bone metabolism and articular cartilage in experimental rats were investigated. Subsequently, the present study assessed dysregulated microRNA (miRNA) in the articular cartilage of experimental rats, and validated the significant miRNA and their targets. Finally, the effects of icariin on chondrocytes in experimental rats and the implicated molecular mechanism were explored. Quantitative polymerase chain reaction demonstrated that icariin significantly reversed DXM-induced bone degradation and stimulated bone regeneration. In addition, some notable changes in articular cartilage were also observed following continuous administration of icariin to DXM-treated rats, including enhanced cartilage area (revealed by safranin-O staining), substantial decrements in serum concentrations of deoxypyridinoline and C-terminal telopeptide of type II collagen, reduced expression of collagen type I and matrix metalloproteinase-13, as well as elevated expression of transforming growth factor-ß. Furthermore, miR-206 was determined to be significantly upregulated in the icariin group compared with the DXM-treated group. A luciferase assay further validated cathepsin K as the target RNA of miR-206. Additionally, icariin (100 µM) facilitated the viability of chondrocytes and reduced apoptotic chondrocytes. More importantly, icariin (100 µM) not only abolished the inhibition effect of DXM on miR-206 expression in chondrocytes, but also eliminated the enhancing effect of DXM on cathepsin K expression. Overall, the present study identified icariin as a novel therapeutic agent in DXM-induced cartilage injury in rats, and revealed that the activation of miR-206 targeting of cathepsin K may be responsible for the chondroprotective efficacy of icariin.

The characteristics of astrocyte on Aß clearance altered in Alzheimer's disease were reversed by anti-inflammatory agent (+)-2-(1-hydroxyl-4-oxocyclohexyl) ethyl caffeate.

Astrocytes are closely related to the amyloid-ß (Aß) deposition in the brain and play crucial roles in Alzheimer's disease (AD) pathology. Meanwhile, inflammation in the CNS has been increasingly demonstrated as a prominent hallmark in AD. Our data from animal models and subjects with Alzheimer's disease (AD) showed GFAP immunoreactivity altered in different stage of AD and had a positive correlation with neprilysin (NEP), suggesting astrocytes might take a protective role in pathogenetic course of AD. Here, we investigate the role of astrocyte in the mechanism of Aß removal. ELISA and western blotting were performed to determine the ability of astrocyte to clear Aß1-42. In this study, we demonstrated that cultured astrocytes removed extracellular oligomeric Aß. However, cultured astrocytes from an AD mouse model showed less capacity to clear extracellular Aß42 but with hyper-expression of NEP protein than normal astrocytes. In addition, LPS-induced inflammation rather than continuous Aß stimuli inhibited the capacity of Aß clearance by astrocytes indicating that inflammation possibly contributed to astrocytic dysfunction. Lastly, HOEC which exhibited anti-inflammatory effects restored the capacity of injured or aged astrocytes to clear Aß. In conclusion, astrocytes have been shown to exert a direct role in Aß clearance and undergo functional impair associated with inflammation in the pathogenesis of AD. Therefore, anti-inflammatory treatments aimed at restoring astrocyte functions may represent an appropriate approach to treat AD.