Ovarian cancer therapy faces significant challenges, including late diagnosis, high recurrence rates, and chemoresistance. The long non-coding RNA GAS5 acts as a tumor suppressor and is downregulated in ovarian cancer, but the mechanism by which it regulates apoptosis remains unclear. This study aimed to reveal the molecular mechanism of GAS5-mediated apoptosis in ovarian cancer cells. We found that GAS5 expression was lower in ovarian cancer tissues and cell lines. Low GAS5 expression was related to worse prognosis. Functional experiments showed that increasing GAS5 expression promoted apoptosis in ovarian cancer cells. Mechanistically, increasing GAS5 raised the mRNA level of the Golgi protein-GOLGA8B. But it reduced the GOLGA8B protein level. Furthermore, GAS5 overexpression shortened the protein half-life and decreased the stability of GOLGA8B, suggesting the involvement of post-transcriptional regulation. Further investigation revealed that GAS5 inhibits O-GlcNAc transferase (OGT), thereby reducing O-GlcNAcylation of GOLGA8B.Ser58 was found to be the key modification site of GOLGA8B. Functionally, GOLGA8B was shown to activate the STAT3 signaling pathway.GAS5 inhibits STAT3 phosphorylation by downregulating GOLGA8B, thereby inducing cell apoptosis. In vivo animal experiments confirmed that GAS5 overexpression suppresses tumor growth. In summary, this study reveals that GAS5 regulates ovarian cancer cell apoptosis by modulating the O-GlcNAcylation of GOLGA8B, which in turn affects the STAT3 signaling pathway, providing a new potential therapeutic target for ovarian cancer.

Accurate identification of antioxidant peptides is crucial for the fields of drug discovery and functional food development. Nevertheless, the effective numerical representation of their variable-length sequences remains challenging. To address this issue, a novel feature-engineering framework is proposed, which is based on logic gates and Huffman coding. The development of two novel descriptors was initiated through the utilization of these methodologies. Subsequently, a robust composite descriptor was formulated by integrating these with classical one-hot encoding. Predictive models constructed on this combined descriptor demonstrated excellent performance in identifying antioxidant peptides. A comparative analysis was conducted, which demonstrated the method's superior accuracy and significant performance enhancement over existing mainstream sequence characterization methods. These results validate the proposed framework as an effective and interpretable approach for deciphering the relationship between peptide sequences and their antioxidant activity.

Alkaline-extracted Platycodon grandiflorum polysaccharide (APGP) exhibits strong immunostimulatory activity, however, the specific structural features and properties that confer the greatest development potential, as well as the underlying mechanism, remain unclear. In this study, the APGP4 fraction exhibiting significant immunostimulatory activity in mouse monocyte-macrophage cells (RAW264.7 cells) and phorbol myristate acetate (PMA)-induced human acute monocytic leukemia cells (THP-1 cells) was identified. This activity was further confirmed through in vivo experiments in immunosuppressed mice. Subsequently, a new polysaccharide, APGP4-3, was isolated and exhibited considerable immunostimulatory activity. APGP4-3 (molecular weight: 11.7 kDa) comprises 1,4-α-D-GalpA, 1,3,4-α-D-Rhap in the backbone, with T-α-L-Araf, 1,3,5-α-L-Araf, 1,3-α-D-Galp, 1,3,4-α-D-Galp and 1,4,6-α-D-Glcp bound to the 3 position of 1,3,4-α-D-Rhap, as well as a few T-α-D-GalpA and Δ4,5-GalpA. Mechanistic studies revealed that the APGP4-3 treatment enhances the proportion of CD11c+ cells (M1-type macrophages) and reduces the proportion of CD206+ cells (M2-type macrophages). Furthermore, APGP4-3 significantly increases the end product (lactic acid) and key enzyme (pyruvate kinase) involved in glycolytic metabolism, while reducing the key enzyme (acetyl coenzyme A) associated with oxidative phosphorylation in macrophages. These findings suggest that APGP4-3 may induce M1 polarization of macrophages by promoting glycolysis and inhibiting oxidative phosphorylation, thereby exerting an immune-promoting effect. This study advances our understanding of the structural features and potential mechanisms underlying the immunostimulatory activity of Platycodon grandiflorum polysaccharide (PGP), and provides new insights into the development of novel immunopotentiators.

Astragalin (AST), a natural flavonoid found in various plants, possesses antioxidant and anti-inflammatory properties. However, its protective efficacy against ultraviolet B (UVB)-induced cutaneous damage remains unclear. This study investigated the photoprotective effects of AST against UVB-induced photodamage using HaCaT keratinocytes and Kunming mice. In vitro, AST mitigated UVB-induced cytotoxicity and apoptosis in HaCaT cells. In vivo, topical application of AST attenuated UVB-induced erythema, epidermal hyperplasia, and collagen degradation in mouse skin. Additionally, AST reduced reactive oxygen species accumulation and enhanced antioxidant enzyme activity via activation of the Keap1/Nrf2 pathway. Furthermore, AST suppressed the expression of proinflammatory cytokines by inhibiting the TLR4/NF-κB signaling pathway. These findings demonstrate the photoprotective properties of AST and support its potential as a natural therapeutic agent for preventing UVB-induced skin damage.

[This retracts the article DOI: 10.3892/br.2018.1084.].

Objective This study examined the antioxidant, whitening, anti-aging, and safety properties of "Sibai" extract obtained from Bombyx Batryticatus, Ampelopsis japonica, Radix Paeoniae Alba, and Atractylodes macrocephala. Methods The "Sibai" extract was optimized by orthogonal design and evaluated for in vitro antioxidant activity through hydroxyl radical scavenging, DPPH scavenging, and total reducing capacity measurements. Tyrosinase inhibition and hemolysis assays assessed its whitening potential and biosafety. Anti-aging effects were examined using Caenorhabditis elegans (C. elegans) lifespan and fecundity assays, while antioxidant capacity under H2O2, juglone, and heat stress was evaluated. Motor function was analyzed via head thrashing, body bending, and spontaneous locomotion tests. Finally, aging-related changes in lipofuscin, reactive oxygen species (ROS), malondialdehyde (MDA), and catalase (CAT) levels were measured. Results The optimal ratio of Bombyx Batryticatus, Ampelopsis japonica, Radix Paeoniae Alba, and Atractylodes macrocephala rhizome was determined to be 3:3:1:1. At specific concentrations, the extract demonstrated significant biological activity: the scavenging rate for hydroxyl radicals was 61.70% (at 48 ​mg/mL), the scavenging rate for DPPH radicals was 93.53% (at 4 ​mg/mL), and its total reducing power at 48 ​mg/mL was comparable to that of Vitamin C (Vc). The tyrosinase inhibition rate was 74.90% (at 24 ​mg/mL), and the hemolysis rate remained below 15%, indicating good in vitro safety. In lifespan experiments using C. elegans as a model, treatment with 48 ​mg/mL extract reduced fecundity and extended the normal lifespan by 38.5%. Under oxidative stress induced by H2O2 and juglone, as well as under heat stress, the survival time of C. elegans in the 48 ​mg/mL treatment group increased by 37.9%, 69.5%, and 49.2% respectively. In terms of motor ability, the 48 ​mg/mL treatment group showed increases in head thrashing and body bending frequencies by 61.4% and 220.7% respectively, along with excellent autonomous movement capacity. At the cellular level, levels of lipofuscin, ROS, and MDA were reduced by 65.8%, 41.5%, and 51.3% respectively, while CAT activity increased by 329.1%. Conclusion This study employed an orthogonal experimental design to determine the optimal formulation. Subsequent validation confirmed that the optimized extract possessed remarkable antioxidant and whitening properties. Furthermore, it was found to significantly enhance stress resistance in C. elegans and improve age-related phenotypes. These results indicated that this formulation may effectively address issues related to aging and oxidative stress.

Breast cancer is one of the most common cancers and the leading cause of cancer-related deaths among women due to the diagnostic delay and failure of treatment.1 Despite the significant progress made in developing therapeutic strategies for breast cancer,effective treatment of breast cancer,particularly aggressive triple-negative breast can-cer,remains lacking.Angiogenesis is a crucial risk factor for breast cancer metastasis and a predictor of poor prognosis.

Legubicin is a novel conjugate of doxorubicin and a legumain-cleavable peptide linker. It has been developed to ameliorate the side effects of doxorubicin. Biodistribution in tumor-bearing mice, acute tolerance, and potential systemic toxic effects in Sprague-Dawley rats and beagle dogs of legubicin were assessed. Legubicin exists mainly as a protein complex in plasma after entering the circulation. Compared with conventional doxorubicin at an equal molar dose in mice, we found higher exposure to doxorubicin in tumor (approximately 1.7-fold increase) while lower exposure in normal tissues (an ~3.26-, 3.46-, and 1.29-fold reduction in heart, kidney, and plasma, respectively) in tumor-bearing mice after intravenous injection of legubicin. The acute maximum tolerance dose (MTD) of legubicin was >16 mg/kg doxorubicin equivalent in female rats, 11 mg/kg doxorubicin equivalent in male rats (LD50 of conventional doxorubicin is 10.51 mg/kg), and >8 mg/kg doxorubicin equivalent in dogs (MTD of conventional doxorubicin is 1.5 mg/kg). Four-week repeat-dose toxicity studies of intravenous legubicin were conducted in rats (5, 10, and 25 mg/kg/dose once weekly) and dogs (3/1.5, 10/5, and 20/10 mg/kg/dose once weekly); the dose levels were reduced from the second dose due to intolerable legubicin-associated toxicity at 20 mg/kg. Major organs of toxicity included the gastrointestinal tract, lymphoid and hematopoietic organs, kidney, skin, liver, reproductive organs, and peripheral nerves, which are all associated with doxorubicin. However, cardiotoxicity was only noted at MTD dose levels. Altogether, our results confirm an improved safety profile of legubicin over conventional doxorubicin and support its clinical benefit for treating cancer.

Oxaliplatin (OXA) is recognized as a first-line drug for gastric cancer. However, low accumulation of the OXA in the target site and the development of drug resistance directly led to treatment failure. In the present study, an ultrasonic extraction method for Atractylodes chinensis (DC.) Koidz. polysaccharides (AKUs) and the combination treatment with OXA in vitro were studied. Results showed that when the pH level was 11, the ultrasound power at 450 W, the solid-liquid ratio was 1:20, and the ultrasound treatment for 30 min, the yield of AKUs was significantly increased to 13.20 +/- 0.35%. The molecular weights of the AKUs ranged from 7.21 to 185.94 kDa, and its monosaccharides were mainly composed of arabinose (Ara), galactose (Gal), and glucose (Glu) with a ratio of 58.36, 16.90, and 15.49%, respectively. Cell experiments showed that, compared to OXA alone (2 mu g/mL, inhibition rate of 18%), the treatment of OXA with AKUs had a significant synergistic inhibitory effect on MKN45 proliferation, which increased to 33, 41, and 45% with increasing AKUs concentrations (5-50 mu g/mL), respectively, representing a 2.5-fold inhibition. Inductively coupled plasma-mass spectrometry (ICP-MS) determination confirmed that AKUs significantly increased the intracellular uptake of OXA by 29%, compared to that of OXA alone. We first demonstrated that the combined synergistic inhibitory effect of AKUs and OXA on gastric cancer cells was mediated by reducing the expression of efflux proteins (MRP1 and MRP2) and increasing the expression of ingested protein (OCT2). As a result of the above, AKUs deserved to be an effective adjuvant combined with chemotherapeutics in a clinical setting.

Lung cancer is a major public health challenge and, despite therapeutic improvements, is the first leading cause of cancer worldwide. The current cure rate from advanced cancer treatment is excessively low. Therefore, it is of great importance to identify novel, potent and less toxic anticancer agents for the treatment of lung cancer. The aim of our research is to synthesize a new biscoumarin 3,3’-((3,4,5-trifluorop -phenyl)methylene)bis(4-hydroxy-2H-chromen-2-one) (C35) as an anticancer agent. C35 was simply prepared by 4-hydroxycoumarin and 3,4,5-trifluorobenzaldehyde under ethanol and its structure was analyzed by spectroscopic analyses. The anti-proliferation effect of C35 was detected using CCK-8 assay. Migration abilities were measured by Transwell assay. The expression of correlated proteins was determined by Western blot. The results showed that C35 displayed strong cytostatic effects on lung cancer cell proliferation. In addition, C35 possessed a significant inhibition of migration by reducing the expression of matrix metalloproteinases-2 (MMP-2) and MMP-9 in lung cancer cells. Furthermore, C35 treatment suppressed the phosphorylation of p38 in lung cancer cells. Moreover, in vivo experiments were carried out, in which we treated Lewis tumor-bearing C57 mice via intraperitoneal injection of C35. Results showed that C35 inhibited tumor growth in vivo. In conclusion, our study demonstrated the anticancer activity of C35 via suppression of lung cancer cell proliferation and migration, which is possibly involved with the inhibition of the p38 pathway.

Malignant insulinoma is an extremely rare type of functioning pancreatic neuroendocrine tumour with a high degree of malignancy and a high incidence of metastasis. However, it is still unclear how malignant insulinomas develop and metastasize. Serum amyloid P component (SAP), a member of the pentraxin protein family, is an acute-phase protein secreted by liver cells. The role of SAP in insulinoma and the related mechanism are still unknown. To determine the effect of SAP on insulinoma, we crossed Rip1-Tag2 mice, which spontaneously develop insulinoma, and SAP knockout (KO) mice to generate Rip1-Tag2;SAP(-/-) mice. We found that SAP deletion significantly promoted the growth, invasion and metastasis of malignant insulinoma through C-X-C motif chemokine ligand 12 (CXCL12) secreted by cancer-associated fibroblasts (CAFs). Further study showed that SAP deletion promoted CXCL12 secretion by CAFs through the CXCR4/p38/ERK signalling pathway. These findings reveal a novel role and mechanism of SAP in malignant insulinoma and provide direct evidence that SAP may be a therapeutic agent for this disease.

Abstract Purpose Transarterial chemoembolization (TACE) with tyrosine kinase inhibitors (TKIs) has been increasingly used to treat unresectable hepatocellular carcinoma (uHCC). However, the superiority of combination therapy to TACE monotherapy remains controversial. Therefore, here we performed a meta-analysis to evaluate the efficacy and safety of TACE plus TKIs in patients with uHCC. Methods We searched four databases for eligible studies. The primary outcome was time to progression (TTP), while the secondary outcomes were overall survival (OS), tumor response rates, and adverse events (AEs). Pooled hazard ratios (HRs) with 95% confidence intervals (95% CIs) were collected for TTP and OS, and the data were analyzed using random-effects meta-analysis models in STATA software. OR and 95% CIs were used to estimate dichotomous variables (complete remission[CR], partial remission[PR], stable disease[SD], progressive disease[PD], objective response rate[ORR], disease control rate[DCR], and AEs) using RStudio’s random-effects model. Quality assessments were performed using the Newcastle–Ottawa scale (NOS) for observational studies and the Cochrane risk of bias tool for randomized controlled trials (RCTs). Results The meta-analysis included 30 studies (9 RCTs, 21 observational studies) with 8246 patients. We judged the risk of bias as low in 44.4% (4/9) of the RCTs and high in 55.6% (5/9) of the RCTs. All observational studies were considered of high quality, with a NOS score of at least 6. Compared with TACE alone or TACE plus placebo, TACE combined with TKIs was superior in prolonging TTP (combined HR 0.72, 95% CI 0.65–0.80), OS (combined HR 0.57, 95% CI 0.49–0.67), and objective response rate (OR 2.13, 95% CI 1.23–3.67) in patients with uHCC. However, TACE plus TKIs caused a higher incidence of AEs, especially hand-foot skin reactions (OR 87.17%, 95%CI 42.88–177.23), diarrhea (OR 18.13%, 95%CI 9.32–35.27), and hypertension (OR 12.24%, 95%CI 5.89–25.42). Conclusions Our meta-analysis found that TACE plus TKIs may be beneficial for patients with uHCC in terms of TTP, OS, and tumor response rates. However, combination therapy is also associated with a significantly increased risk of adverse reactions. Therefore, we must evaluate the clinical benefits and risks of combination therapy. Further well-designed RCTs are needed to confirm our findings. Trial registration PROSPERO registration number: CRD42022298003.

Oxidative stress can lead to various diseases, so achieving antioxidation is crucial for combating these diseases. However, the study of antioxidant mechanism is still in its early stage. Most current methods measure the antioxidant properties of polypeptides using experimental approaches, which are typically time-consuming and laborious. It is therefore urgent to develop a theoretical method to identify antioxidant activity based on peptide sequence information. In this study, amino acid composition, dipeptide composition, and "one-hot" vectors were used to characterize the sequence information of tripeptides. Machine learning methods such as support vector machine, random forest, gradient tree boosting and multi-layer perceptron were utilized to construct models for identifying their antioxidant activities. For the five collected datasets, the Q2 of the random forest model reached 0.864, 0.903, 0.867, 0.611, and 0.703, respectively. Compared with the existing methods 0.061, 0.105, 0.028, 0.121 and 0.111 improvements have been obtained, demonstrating the effectiveness of the current approach. The developed method is expected to significantly contribute to the study of polypeptide antioxidant mechanisms.

For hundreds of years, Atractylodes chinensis (DC.) Koidz. (AK) has been widely used as a treatment for spleen and stomach diseases in China. The AK polysaccharides (AKPs) have been thought to be the important bioactive components. In this stud, the impacts of different extraction methods were analyzed. The differences between AKPs extracted by hot water extraction (HWE), AKPs extracted by ultrasonic extraction (UAE), and AKPs extracted by enzyme extraction (EAE) were compared in terms of yield, total carbohydrate content, molecular weight distribution, monosaccharide composition, and synergistic activity of the AKPs with apatinib were determined. The results indicated that the yield of the polysaccharide obtained from HWE was higher than that of UAE and EAE. However, activity assays indicated that UAE-AKPs and HWE-AKPs enhanced apoptosis of human gastric cancer cells (SGC-7901) treated with apatinib and UAE-AKPs showed the strongest synergistic activities. This is also in agreement with the fact that UAE-AKPs have a smaller molecular weight, β-configuration, and higher galactose content. These findings suggested that UAE is an efficient and environmentally friendly method for producing new polysaccharides from Atractylodes chinensis (DC.) Koidz. for the development of natural synergist and for the treatment of gastric cancer.

Abstract Background Pancreatic cancer (PC) is highly malignant. Chemotherapy is the main treatment strategy, especially for patients with advanced PC. However, chemoresistance has always been a frequently encountered bottleneck. Hence, there is an urgent need to enhance the sensitivity of PC to gemcitabine (GEM). Methods A patient-derived xenograft model was used for screening the hub lncRNAs affecting GEM chemoresistance. Transwell and colony formation assays were used to assess the migration, invasion, and stemness of PC cells. qRT-PCR and western blot analyses were performed to detect the mRNA and protein levels indicated in this study, respectively. The MTT assay was performed to assess the viability of PC cells in vitro. MeRIP-qPCR was used to determine the N6-methyladenosine (m6A) modification level. RIP and pull-down were used to investigate the interaction between SH3BP5-AS1 and miR-139-5p, as well as the interactions among SH3BP5-AS1, ALKBH5, and IGF2BP1. The functional mechanism of SH3BP5-AS1 in PC was identified by gain- and loss-of-function assays and high-throughput sequencing in vitro. Results We demonstrated that SH3BP5-AS1 was significantly upregulated in GEM-resistant PC and predicted a poorer prognosis. SH3BP5-AS1 stability was regulated by ALKBH5/IGF2BP1-mediated m6A modification. Loss of SH3BP5-AS1 reduced PC cell migration and invasion and enhanced the sensitivity of PC to GEM, as confirmed by gain- and loss-of-function assays in vitro and in vivo. Bioinformatics analysis revealed that SH3BP5-AS1 acted as a ceRNA against miR-139-5p and directly targeted CTBP1, affecting the biological behavior of PC cells. The mechanistic studies revealed that the upregulation of SH3BP5-AS1 increased CTBP1 expression by directly activating the Wnt signaling pathway, promoting GEM resistance. Conclusion This study revealed that SH3BP5-AS1 activated Wnt signaling pathway by sponging miR-139-5p, upregulating CTBP1 expression, and contributing to the sensitivity of PC cells to GEM. SH3BP5-AS1 might be a potential target for PC therapy.

[Abstract]: Objective: This study aimed to compare the antibacterial performance of 18 natural plants against six types of microorganisms that tend to grow in cosmetics and select the best natural antiseptic herbs with low toxicity and a broad antibacterial spectrum. The safety performance and effects of the antioxidant, whitening, and reduced inflammation of Coptis chinensis fermentation were explored. Methods: The inhibition performance of the alcoholic extracts for the 18 natural plants against six microorganisms was compared by using the cylinder plate method. The MTT assay was used to determine the cell viability of C. chinensis extract and fermentation products. DPPH, hydroxyl, and ABTS radical scavenging rates as well as iron reduction ability were used to evaluate the antioxidant efficacy. In addition, the activity inhibition rate of tyrosinase and hyaluronidase indicated the whitening and anti-inflammatory properties of C. chinensis extract and fermentation broth. Results: Results showed that among the 18 natural plants, C. chinensis extract had the most prominent inhibition circles and the smallest minimum inhibitory concentration (MIC) value against six microorganisms. After fermentation, the IC50 value of C. chinensis alcoholic extract increased significantly with the cell survival rate, indicating its low biosafety. The free radical scavenging rate, tyrosinase inhibition rate, and hyaluronidase inhibition rate of fermented C. chinensis extract increased significantly, showing better antioxidant activity, whitening activity, and anti-inflammatory activity than alcohol extract. Conclusion: Among the 18 natural plants, C. chinensis has the most substantial antibacterial ability. After fermentation by yeast, the cytotoxicity of C. chinensis decreases, and the antioxidant, whitening, and anti-inflammatory activities increase, indicating that microbial fermentation attenuates toxicity and enhances the effect of plant preservatives.

Abstract Background In our previous study, N end of the Circumsporozoite protein (CSP I-plus) modified recombinant human Endostatin (rEndostatin, endostar) (rES-CSP) was constructed, which had antiangiogenic capability and bound to hepatocellular carcinoma in vivo and in vitro. In this study, the inhibition of rES-CSP on hepatocellular carcinoma metastasis was verified in vivo and in vitro, and its possible mechanism was explored. Methods Firstly, the impact of rES-CSP on the migration, adhesion of hepatoma cell HCCLM3 was identified by wound healing, transwell, and on metastasis of orthotopic xenograft model was identified in nude mouse. Then the expression of metastasis-associated molecules (MMP2, E-cadherin, integrinβ1) and angiogenesis-related factors (VEGFA) in vitro and in vivo were detected by real-time PCR, western blotting, immunohistochemistry. Results Finally, we found that rES-CSP could inhibit the migration and invasion of HCCLM3, and decrease tumor metastasis and growth in nude mouse orthotopic xenograft models. The tumor inhibiting rates of rES-CSP and Endostar were 42.46 ± 5.39% and 11.1 ± 1.88%. The lung metastasis rates of the control, Endostar and rES-CSP were 71, 50, and 42.8%, respectively. Compared with Endostar, rES-CSP significantly down-regulated the expression of VEGFA and integrinβ1. Heparin, a competitive inhibitor of CSP I-plus, which can be bind to the highly-sulfated heparan sulfate proteoglycans (HSPGs) over-expressed in liver and hepatocellular carcinoma, alleviated the down-regulation of VEGFA and integrinβ1. Conclusions These indicate that rES-CSP may play a role in inhibiting tumor growth and metastasis by down-regulating the angiogenic factor VEGF and the metastasis-related molecules or by interfering with HSPGs-mediated tumor metastasis.

Currently, atherosclerosis control is important to prevent future heart attacks or strokes. Protein-enriched extract (PE) from housefly maggots (Musca domestica) can inhibit the development of atherosclerosis partially through its antioxidant effects. Whether PE exerts other anti-atherosclerosis functions remains unclear. Here, PE was found to simultaneously promote cholesterol metabolism effects in apolipoprotein E knockout (ApoE-/- ) mice. Bile acid synthesis plays a key role in regulating cholesterol homeostasis in atherosclerosis. Whether PE alleviates atherosclerosis by promoting bile acid production and consequent cholesterol consumption was further explored. First, 8-week-old male ApoE-/- mice were recruited and fed on a cholesterol-enriched diet. After 8 weeks, these mice were divided into three groups and received gavage administration of PE, simvastatin, and saline for another 8 weeks. Atherosclerosis severity was then assessed. Real-time quantitative polymerase chain reaction and western blot were employed to determine the expression of hepatic ATP-binding cassette transporter A1 (ABCA1), liver X receptor α (LXRα), and peroxisome proliferator-activated receptor-γ (PPAR-γ). Serum levels of high-density lipoprotein-cholesterol (HDL), low-density lipoprotein-cholesterol (LDL), and total cholesterol (TC) were determined by enzyme-linked immunoassay. Results revealed that PE reversed the formation of atherosclerotic lesion; increased the expression of PPAR-γ, LXRα, and ABCA1; increased the amount of bile flow and total bile acid; reduced the serum level of LDL and TC; and increased the level of HDL. In conclusion, enhancement on bile acid production and consequent cholesterol consumption may partially contribute to the anti-atherosclerotic effects of PE. The reversal of PPARγ-LXRα-ABCA1 signaling pathway may be involved in this process.

Platycodon grandiflorum is an edible and medicinal plant, and polysaccharides are one of its important components. To further improve the utilization rate of P. grandiflorum, we investigated the effects of four different extraction methods, including hot water, ultrasonic-assisted, acid-assisted, and alkali-assisted extractions, on the polysaccharides, which were named PG-H, PG-U, PG-C, and PG-A. The findings indicated that the extraction method had a significant impact on the yield, characteristics, and immunoregulatory activity. We observed that the yields decreased in the following order: PG-H, PG-U, PG-C, and PG-A. Galacturonic acid, glucose, galactose, and arabinose were the most prevalent monosaccharides in the four PGs. However, their proportions varied. In addition, the difference between the content of glucose and galacturonic acid was more significant. PG-U had the highest glucose content, whereas PG-C had the lowest. Galacturonic acid content was highest in PG-A, while the lowest in PG-U. The molecular weight decreased in the order of PG-U, PG-H, PG-C, and PG-A; the particle size was in the order of PG-U, PG-A, PG-H, and PG-C. Moreover, the extraction method had a great impact on immunoregulatory activity. The ability to stimulate the immune function of macrophages was as follows: PG-A > PG-C > PG-U > PG-H. The results indicated that PGs, with lower molecular weights and higher GalA content, exhibited better immune-stimulating activity. And more important the AAE method was a good way to extract polysaccharides from Platycodon grandiflorum for use as a functional product and immunological adjuvant.

Background Alternative splicing (AS) of genes has been found to affect gene stability, and its abnormal regulation can lead to tumorigenesis. CELF2 is a vital splicing factor to participate in mRNA alternative splicing. Its downregulation has been confirmed to promote the occurrence and development of pancreatic cancer (PC). However, the regulatory role and mechanisms in PC has not been elucidated. Results CELF2 was downregulated in PC tissues, which affected tumor TNM stage and tumor size, and low expression of CELF2 indicated a poor prognosis of PC. In vivo and in vitro experiments showed that abnormal expression of CELF2 affected the stemness, apoptosis, and proliferation of PC cells. Furthmore, we also found that CELF2 was targeted by ALKBH5 for m6A modification, leading to CELF2 degradation by YTHDF2. Bioinformatic analysis of AS model based on the TCGA database indicated that CELF2 could target CD44 to form different spliceosomes, thereby affecting the biological behavior of PC cells. The conversion of CD44s to CD44V is the key to tumorigenesis. Transcriptomic analysis was conducted to reveal the mechanism of CELF2-mediated CD44 AS in PC. We found that CELF2-mediated splicing of CD44 led to changes in the level of endoplasmic reticulum stress, further regulating the endoplasmic reticulum-associated degradation (ERAD) signaling pathway, thereby affecting apoptosis and cell stemness. In addition, ERAD signaling pathway inhibitor, EerI, could effectively reverse the effect of CD44 on tumors. Conclusions This study indicates that N6-methyladenosine-mediated CELF2 promotes AS of CD44, affecting the ERAD pathway and regulating the biological behavior of PC cells. CELF2 is expected to be a new target for targeted-drug development.