BACKGROUND: The transcription factor POU2F1 regulates the expression levels of microRNAs in neoplasia. However, the miR-29b1/a cluster modulated by POU2F1 in gastric cancer (GC) remains unknown. METHODS: Gene expression in GC cells was evaluated using reverse-transcription polymerase chain reaction (PCR), western blotting, immunohistochemistry, and RNA in situ hybridization. Co-immunoprecipitation was performed to evaluate protein interactions. Transwell migration and invasion assays were performed to investigate the biological behavior of GC cells. MiR-29b1/a cluster promoter analysis and luciferase activity assay for the 3'-UTR study were performed in GC cells. In vivo tumor metastasis was evaluated in nude mice. RESULTS: POU2F1 is overexpressed in GC cell lines and binds to the miR-29b1/a cluster promoter. POU2F1 is upregulated, whereas mature miR-29b-3p and miR-29a-3p are downregulated in GC tissues. POU2F1 promotes GC metastasis by inhibiting miR-29b-3p or miR-29a-3p expression in vitro and in vivo . Furthermore, PIK3R1 and/or PIK3R3 are direct targets of miR-29b-3p and/or miR-29a-3p , and the ectopic expression of PIK3R1 or PIK3R3 reverses the suppressive effect of mature miR-29b-3p and/or miR-29a-3p on GC cell metastasis and invasion. Additionally, the interaction of PIK3R1 with PIK3R3 promotes migration and invasion, and miR-29b-3p , miR-29a-3p , PIK3R1 , and PIK3R3 regulate migration and invasion via the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway in GC cells. In addition, POU2F1 , PIK3R1 , and PIK3R3 expression levels negatively correlated with miR-29b-3p and miR-29a-3p expression levels in GC tissue samples. CONCLUSIONS The POU2F1 - miR-29b-3p / miR-29a-3p-PIK3R1 / PIK3R1 signaling axis regulates tumor progression and may be a promising therapeutic target for GC.
MicroRNAs/metabolism* ; Humans ; Stomach Neoplasms/pathology* ; Cell Line, Tumor ; Cell Movement/physiology* ; Phosphatidylinositol 3-Kinases/metabolism* ; Animals ; Mice ; Octamer Transcription Factor-1/metabolism* ; Mice, Nude ; Class Ia Phosphatidylinositol 3-Kinase/metabolism* ; Neoplasm Invasiveness ; Gene Expression Regulation, Neoplastic/genetics* ; Male ; Immunohistochemistry ; Female

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Eight new diterpenoids, Isodons A-H (1-8), comprising seco-abietane and abietane-type structures, together with 13 known analogues (9-21), were isolated from Isodon lophanthoides (Buch.-Ham. ex D. Don) Hara. The compounds (+)-3/(-)-3, (+)-4/(-)-4, and (+)-5/(-)-5 were identified as three enantiomeric pairs. The planar structures and absolute configurations of 1-8 were determined through high-resolution electrospray ionization mass spectrometry (HR-ESI-MS), 1D & 2D nuclear magnetic resonance (NMR) spectroscopy, electronic circular dichroism (ECD) calculations, and X-ray diffraction crystallography. A cholesterol 7α-hydroxylase (Cyp7a1) luciferase reporter assay revealed significant anti-cholestatic activities for compounds 1, (+)-4, 6, 7, 12-14, and 16. Additionally, compound 6 demonstrated anti-cholestatic effects through the farnesoid X receptor (FXR)-associated signaling pathways in vitro and in vivo. These findings suggest potential applications for I. Lophanthoides in pharmaceutical development.
Abietanes/pharmacology* ; Molecular Structure ; Animals ; Isodon/chemistry* ; Humans ; Diterpenes/pharmacology* ; Plant Extracts/chemistry*

Objective To evaluate the effectiveness of filter membranes made from different materials in monitoring the health status of rodents in barrier environment facilities by investigating their microbial capture performance.Methods Pasteurella pneumotropica(Pp)and Staphylococcus aureus(Sa)were used as representative strains to simulate the process of microbial capture by filter membranes under laboratory conditions.The microbial capture effectiveness of five self-selected filter membranes(M1,M2,M3,M4,and M5)with adsorption and breathability properties and a commercial filter membrane(T1)were comprehensively evaluated based on captured dust mass,minimum detection limit,and differences in Ct values obtained through fluorescence quantitative PCR detection.The best-performing self-selected filter membrane was placed in the ventilation ducts of cage racks within the barrier facility,with sentinel mice in corresponding cage racks as the control group.Staphylococcus epidermidis and Escherichia coli were used as indicator bacteria to calculate the positive detection rate and coincidence rate,thereby exploring the feasibility of using microbial capture filter membranes to monitor the health status of experimental animals in barrier facilities.Results In terms of the captured dust mass,the self-selected filter membrane M3(non-woven filter membrane with a diameter of 0.1 μm);showed a capture effectiveness second only to T1,with a capture mass of 0.126 g.For Sa,all filter membranes except M4 had a minimum detection limit of 102 CFU/g.For Pp,the minimum detection limit for all filter membranes was 102 CFU/g.However,the Ct value of the quantitative fluorescence PCR amplification results for M3 was significantly lower than that of other materials,indicating that M3 had the best capture performance among the five self-selected materials.In the filter detection verification experiment,the positive detection rate of Staphylococcus epidermidis in sentinel mouse feces and M3 was 50.00％(6/12)and 58.33％(7/12),respectively,with a coincidence rate of 92％.The positive detection rate of Escherichia coli in both sentinel mouse feces and M3 was 50.00％(6/12),with a coincidence rate of 100％.Conclusion Among the 5 self-selected filter membranes,M3 exhibits the best capturing performance.Within the barrier environment facilities,M3 outperforms sentinel mice in monitoring Staphylococcus epidermidis.Therefore,non-woven filter membrane with a diameter of 0.1 μm;can be used as the material for microbial capture filter membranes,providing valuable insights for the selection and application of microbial capture filter membranes used in PCR monitoring of cage exhaust air dust.

Wound healing is a complex process of biological integration in which the adverse conditions such as excessive inflammatory reactions, cell proliferation and migration disorders, and cellular secretion impairment can lead to refractory wounds. Characterized by complex etiology, protracted condition, and high morbidity and recurrence rate, refractory wounds severely impair patients′ physical and mental health. In clinical practice, refractory wounds are primarily treated with surgical debridement and skin transplantation, but there still exist problems such as large surgical wounds, prolonged recovery time, and high recurrence rate. In recent years, owing to the multipotent differentiation, immunomodulatory, and paracrine functions of stem cells, cell-assisted lipotransfer (CAL) technique, which involves intra-body injection of a mixture of autologous adipose-derived stem cells (ADSCs) and granular fat for refractory wound repair, has demonstrated promising application prospects. It is of great significance in its clinical application to clarify the mechanism of effects of CAL technique in refractory wound repair. The authors reviewed the research progress in the mechanism of effects of CAL technique in repairing refractory wounds to provide references for the research and treatment of refractory wounds.

Purpose: Mixed-lineage leukemia protein 4 (MLL4/KMT2D) is a histone methyltransferase, and its mutation has been reported to be associated with a poor prognosis in many cancers, including lung cancer. We investigated the function of MLL4 in lung carcinogenesis. Materials and Methods: RNA sequencing (RNA-seq) in A549 cells transfected with control siRNA or MLL4 siRNA was performed. Also, we used EdU incorporation assay, colony formation assays, growth curve analysis, transwell invasion assays, immunohistochemical staining, and in vivo bioluminescence assay to investigate the function of MLL4 in lung carcinogenesis. Results: We found that MLL4 expression was downregulated in non–small cell lung cancer (NSCLC) tissues compared to adjacent normal tissues and tended to decrease with disease stage progression. We analyzed the transcriptomes in control and MLL4- deficient cells using high-throughput RNA deep sequencing (RNA-seq) and identified a cohort of target genes, such as SOX2, ATF1, FOXP4, PIK3IP1, SIRT4, TENT5B, and LFNG, some of which are related to proliferation and metastasis. Our results showed that low expression of MLL4 promotes NSCLC cell proliferation and metastasis and is required for the maintenance of NSCLC stem cell properties. Conclusion Our findings identify an important role of MLL4 in lung carcinogenesis through transcriptional regulation of PIK3IP1, affecting the PI3K/AKT/SOX2 axis, and suggest that MLL4 could be a potential prognostic indicator and target for NSCLC therapy.

Objective: To investigate the effect and the mechanism of Astragalus membranaceus (Huangqi in Chinese, HQ) extract on the intestinal absorption of six alkaloids of Aconitum carmichaelii (Fuzi in Chinese, FZ) in rats with spleen deficiency and provide novel insights into the application of HQ on modulating intestinal barrier. Methods: Four-week-old male Sprague-Dawley rats were fed with Xiaochengqi Decoction to induce the spleen deficiency model for 40 d. Single-pass intestinal perfusion model were used to study the effects of HQ extract on the absorption of alkaloids. Protein expression and mRNA levels of MRP2 and BCRP and tight junction proteins (TJ, including Claudin-1, Occludin and ZO-1) were measured using Western blot and real-time PCR, respectively. The location and expression of TJ protein was also investigated by the immunofluorescence method. Results: Compared with the normal group, the protein expression of MRP2, BCRP and TJ proteins in the model group were significantly down-regulated. After oral administration of HQ, the alkaloid absorption in intestinal villi was inhibited, MRP2, BCRP and TJ proteins were up-regulated, the green fluorescence staining of Claudin-1, Occludin, and ZO-1 was enhanced, and a thick layer of mucus was deposited on the surface of the epithelium of the intestinal cavity. Conclusion: HQ as an intestinal barrier modulator improves the physiological changes of the intestinal environment of spleen deficiency to reduce the absorption of toxic components, leading to a decrease in the absorption of drug-like molecules.

Hepatocellular carcinoma (HCC) is the most common primary liver malignancy and is the fourth-leading cause of cancer-related deaths worldwide. HCC is refractory to many standard cancer treatments and the prognosis is often poor, highlighting a pressing need to identify biomarkers of aggressiveness and potential targets for future treatments. Kinesin family member 2C (KIF2C) is reported to be highly expressed in several human tumors. Nevertheless, the molecular mechanisms underlying the role of KIF2C in tumor development and progression have not been investigated. In this study, we found that KIF2C expression was significantly upregulated in HCC, and that KIF2C up-regulation was associated with a poor prognosis. Utilizing both gain and loss of function assays, we showed that KIF2C promoted HCC cell proliferation, migration, invasion, and metastasis both in vitro and in vivo. Mechanistically, we identified TBC1D7 as a binding partner of KIF2C, and this interaction disrupts the formation of the TSC complex, resulting in the enhancement of mammalian target of rapamycin complex1 (mTORC1) signal transduction. Additionally, we found that KIF2C is a direct target of the Wnt/β-catenin pathway, and acts as a key factor in mediating the crosstalk between Wnt/β-catenin and mTORC1 signaling. Thus, the results of our study establish a link between Wnt/β-catenin and mTORC1 signaling, which highlights the potential of KIF2C as a therapeutic target for the treatment of HCC.
Adult ; Aged ; Animals ; Carcinoma, Hepatocellular/pathology* ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Epithelial-Mesenchymal Transition/genetics* ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Intracellular Signaling Peptides and Proteins/metabolism* ; Kinesins/metabolism* ; Liver Neoplasms/pathology* ; Male ; Mice ; Mice, Inbred BALB C ; Middle Aged ; Neoplasm Staging ; Prognosis ; Protein Binding ; RNA, Small Interfering/metabolism* ; Survival Analysis ; Tumor Burden ; Wnt Signaling Pathway ; Xenograft Model Antitumor Assays ; beta Catenin/metabolism*

[Abstract ] Objective More and more researches demonstrate that epithelial-mesenchymal transition(EMT) is highly rele-vant to cancer metastasis , tissue fibrosis and skin wound healing , but its role in the wound healing of oral mucosa still needs further ex-ploration.This study was designed to discuss the effects of EMT on the wound healing of lingual mucous membrane in rats . Methods A 3mm-diameter circular defect was made on the mucosa of lingual dorsum in 8-week-old Wistar rats.The rats were put to death at day 0, 1, 2, 3, 4 and 8 after operation.We observed tissue healing process by HE staining and used immunohistochemical fluorescence to detect the expressions of EMT-related protein E-cadherin(E-cad), Vimentin and Fibroblast specific proteins (FSP1) in lingual mucous membrane during wound healing . Results The expressions of E-cad at the edge of the wound in the lingual mucous membrane of rats at day 2 and 4 after operation were less than those at day 0 and 1 after operation , while interstitial cell specific protein Vimentin and FSP1 were positively expressed in the epidermal basal layer .The wound of lingual mucous membrane healed completely at day 8 after operation , and E-cad expression was close to that at day 0 after operation .A small amount of Vimentin and FSP 1 expressed in the epi-thelial basal layer . Conclusion E-cad, Vimentin, FSP1 are involved in the wound healing process of lingual mucous membrane . During the wound healing in the tongue mucous membrane of rats , some epithelial cells gain the feature of mesenchymal cells in the process of migration to wound centre .EMT has played an important role during wound healing of lingual mucous membrane in rats .