Punicalagin, a major ellagitannin enriched in pomegranate (Punica granatum L.) peel, has been implicated in cellular redox regulation, yet its integrated mechanism against oxidative hepatocellular injury remains insufficiently defined. Here, we combined network pharmacology, molecular docking, and in vitro validation to elucidate the protective actions of punicalagin against hydrogen peroxide (H2O2)-induced injury in HepG2 cells. Punicalagin pretreatment markedly improved cell viability and suppressed intracellular reactive oxygen species accumulation following oxidative challenge. This protection was associated with improved mitochondrial function. Flow cytometry further demonstrated that punicalagin attenuated H2O2-triggered apoptosis, consistent with modulation of the mitochondrial apoptotic cascade as evidenced by an increased Bcl-2/Bax balance and reduced caspase-3 activation. Punicalagin also enhanced autophagy, as indicated by increased LC3B-II and decreased p62. Network pharmacology identified overlapping targets between punicalagin and liver injury, with enrichment analyses highlighting PI3K/AKT-related signaling; docking predicted favorable binding of punicalagin to AKT1. Mechanistically, punicalagin blunted oxidative stress-induced phosphorylation of PI3K, AKT, and mTOR without altering total protein abundance. Importantly, pharmacological reactivation of AKT using SC79 mitigated punicalagin-induced autophagy, supporting a pathway-dependent mode of action. Collectively, these findings indicate that punicalagin alleviates oxidative hepatocellular injury, supporting its potential as a food-derived bioactive for oxidative stress-related liver dysfunction.

BACKGROUND:Diabetes mellitus (DM) and metabolic-associated fatty liver disease (MAFLD) are common metabolic disorders, and their coexistence can exacerbate the progression of either disease. Human umbilical cord mesenchymal stem cell (hUC-MSC) therapy has shown promising potential in the treatment of several metabolic diseases. AIM:To investigate how hUC-MSCs affect liver metabolism in diabetic rats with MAFLD and assess their therapeutic potential and underlying mechanisms. METHODS:A streptozotocin-induced rat model of DM with MAFLD was established, and hUC-MSCs were administered via tail vein injection. Changes in body weight, fasting blood glucose (FBG), and serum triglyceride (TG), alanine aminotransferase, aspartate aminotransferase levels, and pathological changes of liver were evaluated. Receiver operating characteristic analysis was used to assess the diagnostic value of differential metabolites and their ability to predict the therapeutic effects of hUC-MSCs. Spearman correlation was employed to analyze the relationships between liver metabolites and key biochemical markers. RESULTS:hUC-MSC treatment significantly reduced FBG and TG levels in diabetic rats with MAFLD and improved histological steatosis and injury in the liver. Metabolomic analysis indicated that hUC-MSCs significantly ameliorated liver metabolic disturbances via their regulatory effect on several key metabolic pathways related to carbohydrate, amino acid, and lipid metabolism. Receiver operating characteristic curve analysis revealed that 70 differential metabolites had good diagnostic value for DM with MAFLD and could effectively predict the therapeutic effect of hUC-MSCs. Moreover, Spearman correlation analysis confirmed that significant correlations existed between differential liver metabolites and the concentrations of biochemical markers (FBG, TG, alanine aminotransferase, aspartate aminotransferase). CONCLUSION:hUC-MSCs alleviate liver metabolic disturbances in diabetic rats with MAFLD, thereby mitigating the pathological state of DM and slowing the progression of MAFLD.

Nephrotic syndrome (NS) is a relatively rare and serious presentation of IgA nephropathy (IgAN) (NS-IgAN). Previous research has suggested that the pathogenesis of NS-IgAN may involve circulating immune imbalance and kidney injury; however, this has yet to be fully elucidated. To investigate the cellular and molecular status of NS-IgAN, we performed single-cell RNA sequencing (scRNA-seq) of peripheral blood mononuclear cells (PBMCs) and kidney cells from pediatric patients diagnosed with NS-IgAN by renal biopsy. Consistently, the proportion of intermediate monocytes (IMs) in NS-IgAN patients was higher than in healthy controls. Furthermore, flow cytometry confirmed that IMs were significantly increased in pediatric patients with NS. The characteristic expression of VSIG4 and MHC class II molecules and an increase in oxidative phosphorylation may be important features of IMs in NS-IgAN. Notably, we found that the expression level of CCR2 was significantly increased in the CMs, IMs, and NCMs of patients with NS-IgAN. This may be related to kidney injury. Regulatory T cells (Tregs) are classified into two subsets of cells: Treg1 (CCR7high, TCF7high, and HLA-DRlow) and Treg2 (CCR7low, TCF7low, and HLA-DRhigh). We found that the levels of Treg2 cells expressed significant levels of CCR4 and GATA3, which may be related to the recovery of kidney injury. The state of NS in patients was closely related to podocyte injury. The expression levels of CCL2, PRSS23, and genes related to epithelial-mesenchymal transition were significantly increased in podocytes from NS-IgAN patients. These represent key features of podocyte injury. Our analysis suggests that PTGDS is significantly downregulated following injury and may represent a new marker for podocytes. In this study, we systematically analyzed molecular events in the circulatory system and kidney tissue of pediatric patients with NS-IgAN, which provides new insights for targeted therapy in the future.

Imaging the kidney is necessary to evaluate its function and track the progression of renal diseases, but it remains difficult due to invasive damage, radiation, high cost, etc. Fluorescence imaging (FLI) has received increasing attention due to its benefits, and Pillar[n]arene has attracted widespread interest due to its rigid structure, facile synthesis, and ease of functionalisation, demonstrating great imaging potential. Therefore, a novel type of functionalised Pillar[5]arene (P5DMP) with 10 positive charges was designed and created. P5DMP is a potential fluorescent material due to its major advantages, including stability, water solubility, strong biocompatibility, and superior fluorescence qualities. Notably, the positive charge design of P5DMP allows it to collect in the negatively charged filtration barrier to achieve focused accumulation and imaging of the kidney. In fact, the in vivo experiments indicated that P5DMP accumulated preferentially in the kidney with the greatest distribution in the glomeruli. The findings of the present study indicated that P5DMP may achieve a targeted and precise image of the kidney and may provide a simple and universal method for the fabrication of an outstanding renal imaging agent with significant clinical application potential.

HCC is one of the most common malignant tumors. The life and health of humans are gravely threatened by HCC because of its hidden onset, high recurrence rate, poor therapeutic effect, and high mortality. It is essential to explore the particular pathological mechanisms of HCC in order to increase the rate of early diagnosis and enhance patient therapy outcomes. Recent research has demonstrated that SCUBE3 can influence HCC cell proliferation by regulating the TGFβ/PI3K/AKT/GSK3β pathway. The molecular regulatory network of HCC proliferation is improved by this research, which also offers a solid theoretical and experimental foundation for SCUBE3 as a potential new therapeutic target for HCC.

Subsequently to the publication of the above article, an interested reader drew to the authors' attention that two pairs of data panels in Fig. 7D on p. 1008, showing the results from Transwell invasion assay experiments, contained overlapping sections such that these panels were likely to have been derived from the same original sources where they were intended to show the results from differently performed experiments. After having consulted their original data, the authors were able to identify that two of the data panels in Fig. 7D were inadvertently selected incorrectly; specifically, the 'GST+SB203580' and 'GST‑hS100A9+PD98059' panels in this figure. The revised version of Fig. 7, showing the correct data panels for the 'GST+SB203580' and 'GST‑hS100A9+PD98059' panels in Fig. 7D, is shown on the next page. The authors confirm that the errors made during the assembly of Fig. 7 did not grossly affect the major conclusions presented in this paper, and are grateful to the Editor of International Journal of Oncology for allowing them this opportunity to publish a Corrigendum. They also apologize to the readership for any inconvenience caused. [International Journal of Oncology 42: 1001-1010, 2013; DOI: 10.3892/ijo.2013.1796].

An accurate diagnosis of acute kidney injury (AKI) at the early stage is critical to not only allow preventative treatments in time but also forecast probable medication toxicity for preventing AKI from starting and progressing to severe kidney damage and death. Therefore, supramolecular fluorescent biomaterials based on Q [8] and PEG-APTS have been prepared herein. This study has found that the unique properties of outer surface methine and the positive density of Q [8] can form a stable assembly with PEG-APTS, and has provided the possibility for the faster crossing of the glomerular filtration barrier to enter into the resident cells of the kidney. In addition to the excellent fluorescence properties, the as-synthesized biomaterial Q [8]@PEG-APTS has possessed significantly low biological toxicity. Most importantly, the accumulation of Q [8]@PEG-APTS in large amounts in cytoplasm and nucleus of HK2 and HMCs cells, respectively, within 24 h enabled distinguishing kidney cells when diagnosing and providing some foundation for early AKI.

Periodontitis is a chronic infectious disease that alters the cellular microenvironment and promotes bone absorption. Bone morphogenetic protein 9 (BMP9) serves an important role in proliferation and differentiation, and tumor necrosis factor‑alpha (TNF‑α) is an important contributor to bone resorption. The present study aimed to investigate the effect of osteogenic differentiation in the presence of BMP9 and TNF‑α in rat follicle stem cells (rDFCs). rDFCs were transfected with adenoviruses expressing BMP9 (AdBMP9) and the expression levels of important proteins [BMP9, β‑catenin, glycogen synthase kinase 3β (GSK3β), phosphorylated‑GSK3β, calcium/calmodulin dependent protein kinase II and nemo like kinase] were determined using western blotting. The effect of osteogenesis was analyzed using reverse transcription‑quantitative PCR, in addition to alkaline phosphatase, Alizarin Red S, and hematoxylin and eosin staining methods. The results of the present study revealed that TNF‑α activated the canonical Wnt signaling pathway and suppressed osteogenesis. High concentrations of Dickkopf 1 (DKK1) reduced the osteogenic differentiation of AdBMP9‑transduced rDFCs, whereas low concentrations of DKK1 promoted BMP9‑induced bone formation, which was discovered to partially act via the canonical and non‑canonical Wnt signaling pathways. In conclusion, the findings of the present study suggested that the enhanced promoting effect of BMP9 alongside the treatment with low concentrations of DKK1 may be useful for treating periodontitis bone absorption.

目的 探索骨形成蛋白9(bone morphogenetic protein 9,BMP9)在诱导大鼠牙囊干细胞(rat dental follicle stem cells,rDFCs)成骨向分化过程中的Smad信号通路调控机制.方法 重组腺病毒骨形成蛋白9 (recombinant adenoviruses expressing BMP9,Ad-BMP9)转染纯化的第3代rDFCs后,Realtime qPCR、碱性磷酸酶(alkaline phosphatase,ALP)染色及活性检测观察BMP9调节rDFCs早期及中晚期成骨能力,茜素红S染色检测钙盐沉积,Western blot检测Smad1/5/8磷酸化水平.结果 BMP9促进rDFCs成骨因子表达:成骨转录因子Runx2、成骨细胞特异性转录因子Osterix、ALP及骨桥蛋白(osteopontin,OPN)在BMP9刺激组较空白组均明显升高(P＜0.05),钙盐沉积及钙结节形成也明显多于空白组,且BMP9促进了Smad1/5/8蛋白磷酸化水平升高.结论 BMP9通过激活Smad信号通路,提高Smad1/5/8蛋白磷酸化水平从而促进早中晚期成骨因子及碱性磷酸酶表达,促进了rDFCs成骨向分化.

Interleukin 6 (IL-6), a tumor promoting cytokine, has been largely implicated in the development of renal cell carcinoma (RCC). Hepatocyte cell adhesion molecule (hepaCAM) is a novel tumor suppressor, which is lost or down-regulated in many cancer types including RCC. In the present study, we intensively investigated the connection between IL-6 and hepaCAM in RCC. Our analysis of RCC tissues, adjacent tissues and paired serum samples from RCC patients revealed that IL-6 was elevated in patient serum and RCC tissue, whereas hepaCAM was completely lost or significantly down-regulated. Furthermore, we observed an association between IL-6 increase and hepaCAM decrease in RCC tissue samples. In the section of cytological researches, we found in RCC cell lines that IL-6 was a direct upstream regulator of hepaCAM, and that hepaCAM down-regulation was involved in IL-6-driven cell proliferation. We also demonstrated that IL-6-mediated promoter hypermethylation largely accounted for the hepaCAM loss in RCC, and it was STAT3-dependent. Additionally, our data showed that DNMT1 up-regulation induced by IL-6/STAT3 signaling was indispensable for IL-6-mediated hepaCAM loss in RCC cell lines ACHN and 769-P, while DNMT3b up-regulation was crucial for hepaCAM loss in A498. Our findings provide a novel signal pathway regulating cell proliferation, potentially representing a therapeutic target for RCC.

Cells, scaffolds, and growth factors play important roles in bone regeneration. Bone morphogenetic protein 9 (BMP9), a member of BMP family, could facilitate osteogenesis by regulating growth factors and promoting angiogenesis. Similar to other stem cells, rat dental follicle stem cells (rDFCs), the precursor cells of cementoblasts, osteoblasts and periodontal ligament cells, can self-renew and exhibit multipotential capacity. Coralline hydroxyapatite (CHA) has good biocompatibility and conductivity required for bone tissue engineering. Here, we reported that BMP9 could enhance the osteogenic differentiation of rDFCs in cell culture. Moreover, our results suggested that BMP9 acted through the Smad1/5/8 signaling pathway. We also produced a novel scaffold that encompasses bio-degradable CHA seeded with recombinant adenoviruses expressing BMP9-transfected rDFCs (Ad-BMP9-transfected rDFCs). With this implant, we achieved more alveolar bone regeneration in the alveolar bone defect compared to blank group, CHA group and rDFCs group. Our results provided a novel bio-implants composed of Ad-BMP9-transfected rDFCs and CHA scaffolds and its mechanism is regarding the activation of Smad1/5/8 signaling pathway in BMP9-induced rDFCs osteogenesis.

Periodontal ligament stem cells (PDLSCs) with bone morphogenic ability are used to treat diseases such as periodontitis. Their treatment potential is increased when used in combination with proteins that induce osteogenic differentiation. For example, bone morphogenetic protein-9 (BMP9) has been found to have potent osteogenic activity. In the present study, PDLSCs were isolated from human periodontal membrane and infected with recombinant adenoviruses expressing BMP9 (Ad-BMP9). Levels of osteogenic markers such as runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), osteopontin (OPN), and osteocalcin (OCN) as well as mineralization ability were measured. The results showed that BMP9 promoted bone formation of PDLSCs. In other experiments, SB203580 and PD98059, which are inhibitors of p38 and ERK1/2, respectively, were used to determine if these kinases are involved in the osteogenic differentiation process. The resulting protein expression profiles and osteogenic markers of PDLSCs revealed that the mitogen-activated protein kinase (MAPK) signaling pathway might play an important role in the process of BMP9-induced osteogenic differentiation of PDLSCs.

Dental follicle stem cells are a group of cells possessing osteogenic, adipogenetic and neurogenic differentiations, but the specific mechanism underlying the multilineage differentiation remains still unclear.Great attention has been paid to bone morphogenetic protein-9 (BMP-9) due to its potent osteogenic activity.In the present study, rat dental follicle stem cells were isolated and purified, and cells of passage 3 underwent adenovirus mediated BMP-9 gene transfection to prepare dental follicle stem cells with stable BMP-9 expression.Detection of alkaline phosphatase (ALP) and calcium deposition showed dental follicle stem cells transfected with BMP-9 gene could significantly promote the osteogenesis.In addition, SB203580 and PD98059 were employed to block the p38 mitogen-activated protein kinase (p38MAPK) and extracellular signal-regulated kinase (ERK1/2), respectively.Detection of ALP and calcium deposition revealed the BMP-9 induced osteogenic differentiation of dental follicle stem cells depended on MAPK signaling pathway.

Background: Preterm premature rupture of the membrane is a frequent complication during gestational period. Three-dimensional culture can help damaged tissue restore anatomical integrality, being worth for clinical application. Objective: To explore biological characteristics of human amniotic cells three-dimensionally cultured by fibrin scaffold, study the possibility of cell/scaffold composite material for repairing preterm premature rupture of the membrane, and to compare with normal monolayer culture. Design, Time and Setting: An in vitro cell/scaffold study was performed at the Basic Institute of Chongqing University of Medical Science between December 2007 and November 2008. Materials: Amnion tissue derived from uterine-incision delivery women was provided by the Department of Obstetrics and Gynecology, the First Affiliated Hospital, Chongqing University of Medical Sciences; fibrinogen was provided by Sigma, USA. Methods: Either epithelial cells or mesenchymal cells, being isolated from human amniotic membrane, were cultivated using enzyme digestion and repeated adherence methods. Fibrinogen was conglomerated, and epithelial cells of logarithmic growth were plated onto the surface of fibrin scaffold to simulate a three-dimensional culture in vitro. Before conglomerating fibrinogen, mesenchymal cells of logarithmic growth were mixed with fibrin solution to simulate a three-dimensional culture in vivo. In addition, either epithelial cells or mesenchymal cells were incubated onto 24-well plate for normal monolayer culture. Main Outcome Measures: Cell morphology using inverted microscopy, proliferation of mesenchymal cells, and interaction between mesenchymal cells and scaffold under different culture environments. Results: Epithelial cells were round and smooth on the surface of fibrin scaffold, pseudopodia were stretched out, and microvillus were rich. Mesenchymal cells in fibrin glue were fusiform in shape and stretched out along scaffold. Cells formed net structure at different surfaces. By three-dimensional culture, proliferation of mesenchymal cells was stable but slow compared to Monolayer culture. By in vivo three-dimensional culture, fibrin glue gradually shrank and gel thickness gradually decreased. On the 5th day, the gel thickness was only 40% for the initial thickness, and on the 15th day, gel thickness was 10%. Conclusion: Both epithelial cells and mesenchymal cells can sterically grow on fibrin scaffold, and the proliferation is stable. Fibrin glue gradually shrinks in cell/fibrin composite scaffold, suggesting that amniotic cells/fibrin composite scaffold can simulate body tissues to repair preterm premature rupture of the membrane.