IL-12 improves the anti-HCC efficacy of dendritic cells loaded with exosomes from overexpressing Rab27a tumor cells.

Determining the appropriate source of antigens for optimal antigen presentation to T cells is a major challenge in designing dendritic cell (DC) -based therapeutic strategies against hepatocellular carcinoma (HCC). Tumor-derived exosomes (Tex) express a wide range of tumor antigens, making them a promising source of antigens for DC vaccines. As reported, the exosomes secreted by tumor cells can inhibit the antitumor function of immune cells. In this study, we transfected hepatocellular carcinoma cells with Rab27a to enhance the yield of exosomes, which were characterized using transmission electron microscopy and Western blot analysis. We found that Tex secreted by overexpressing Rab27a Hepatocellular carcinoma cell lines pulsed DC is beneficial for the differentiation and maturation of DCs but inhibits the secretion of the IL-12 cytokine. Consequently, we developed a complementary immunotherapy approach by using Tex as an antigen loaded onto DCs, in combination with the cytokine IL-12 to induce antigen-specific cytotoxic T lymphocytes （CTLs）. The results indicated that the combination of DC-Tex and IL-12 was more effective in stimulating T lymphocyte proliferation, releasing IFN-γ， and enhancing cytotoxicity compared to using exosomes or IL-12 alone. Additionally, the inclusion of IL-12 also compensated for the reduced IL-2 secretion by DCs caused by Tex. Moreover, in a BALB/c nude mice model of hepatocellular carcinoma, CTLs induced by DC-Tex combined with IL-12 maximized the tumor-specific T-cell immune effect and suppressed tumor growth. Thus, Tex provides a novel and promising source of antigens, with cytokines compensating for the shortcomings of Tex as a tumor antigen. This work helps to clarify the role of exosomes in tumor immunotherapy and may offer a safe and effective prospective strategy for the clinical application of exosome-based cellular immunotherapy.

Fermentation of waste water from agar processing with Bacillus subtilis by metabolomic analysis.

Fungal infection has become a major threat to crop loss and affects food safety. The waste water from agar processing industries extraction has a number of active substances, which could be further transformed by microorganisms to synthesize antifungal active substances. In this study, Bacillus subtilis was used to ferment the waste water from agar processing industries extraction to analyze the antifungal activity of the fermentation broth on Alternaria alternata and Alternaria spp. Results showed that 25% of the fermentation broth was the most effective in inhibited A. alternata and Alternaria spp., with fungal inhibition rates of 99.9% and 96.1%, respectively, and a minimum inhibitory concentration (MIC) was 0.156 µg/mL. Metabolomic analysis showed that flavonoid polyphenols such as coniferyl aldehyde, glycycoumarin, glycitin, and procyanidin A1 may enhance the inhibitory activity against the two pathogenic fungal strains. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that polyphenols involved in the biosynthesis pathways of isoflavonoid and phenylpropanoid were upregulated after fermentation. The laser confocal microscopy analyses and cell conductivity showed that the cytoplasm of fungi treated with fermentation broth was destroyed. This study provides a research basis for the development of new natural antifungal agents and rational use of seaweed agar waste. KEY POINTS: • Bacillus subtilis fermented waste water has antifungal activity • Bacillus subtilis could transform active substances in waste water • Waste water is a potential raw material for producing antifungal agents.

Down-regulation of miR-424 inhibited the metastasis of endometrial carcinoma via targeting PTEN/PI3K/AKT signaling pathway.

Background: The incidence of endometrial carcinoma (EC) has been increasing annually, and treatment of advanced cases remains challenging. MicroRNA-424 (miR-424) was reported to affect several types of tumors, but its role in EC has not been studied. Methods: We generated transient knockdown models of miR-424 and PTEN in EC cells. We measured mRNA and protein expression using RT-PCR and western blotting. We evaluated cell proliferation, invasion, migration, and apoptosis using CCK8, Transwell, wound healing, and flow cytometry assays. We also investigated the effect of miR-424 and PTEN on tumor growth using a metastatic tumor model in nude mice. Results: The expression of miR-424 was significantly elevated in EC tissues and cell lines. MiR-424 inhibitor significantly restrained PTEN/PI3K/AKT signaling, while miR-424 mimic activated this pathway. Knockdown of PTEN significantly reversed the effects of miR-424 inhibitor on cell proliferation, invasion, migration, and apoptosis in EC cells. The significant inhibition of tumor growth and ki67 expression caused by miR-424 inhibitor were markedly promoted by sh-PTEN. Conclusions: Our findings suggest that miR-424 inhibitor could inhibit cell proliferation, invasion, migration, epithelial-mesenchymal transition (EMT) process, and tumor growth, while promoting apoptosis in EC. However, the effects of miR-424 inhibitor were markedly reversed by sh-PTEN. This study provides a potential novel therapeutic strategy for the prevention and treatment of EC by targeting miR-424.

Assessing Metabolic Risk Factors for LVSI in Endometrial Cancer: A Cross-Sectional Study.

Objective: This study analyzed metabolic factors associated with lymphovascular space invasion (LVSI) and compared the difference between type 1 and type 2 endometrial cancer (EC). Methods: Four hundred patients primarily diagnosed with EC who underwent hysterectomy with pathological results at Fujian Medical University Cancer Hospital from January 2019 to January 2021 were included. Demographic variable data were collected as well as pathological results. Laboratory evaluations included fasting blood glucose (FBG), serum cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL), apolipoprotein A (Apo A) and apolipoprotein B (Apo B). Characterization of binary logistic regression models was used to test the odds ratios (ORs) between LVSI and its metabolic parameters with different subtypes of EC. Results: The results indicated that CA125, ROMA, Ki67 score, FBG and TC were higher in EC patients with LVSI (all p<0.05). Negative ER and PR expression was positively associated with LVSI (P<0.05). CA125, ROMA, FBG, TC and ER were found to be independent risk factors for LVSI. CA125, ROMA and FBG were significantly elevated in type 1 EC patients with LVSI compared with those without LVSI (all p<0.05). TC and Ki67 scores were much higher in type 2 EC patients with vs without LVSI (all p<0.05). Negative PR expression was positively related to both type 1 and type 2 EC patients with LVSI. Consequently, CA125, ROMA, FBG and Apo B were found to be independent risk factors for LVSI in type 1 EC, and TC was found to be an independent risk factor for LVSI in type 2 EC. Conclusion: FBG and TC were both independent risk factors for LVSI in EC. FBG and Apo B were independent risk factors for LVSI in type 1 EC. TC was an independent risk factor for LVSI in type 2 EC.

Identification of exosomal hsa-miR-483-5p as a potential biomarker for hepatocellular carcinoma via microRNA expression profiling of tumor-derived exosomes.

To date, most studies of exosomes related to hepatocellular carcinoma (HCC) have used commercial cancer cell lines or patient plasma as source material. In this study, we isolated exosomes directly from HCC tissues to investigate the potential of exosomal contents as biomarkers for HCC. Exosomes were identified and verified using transmission electron microscopy, nano-flow cytometry analysis, and western blotting. Tissue-derived exosomal miRNA expression was profiled by high-throughput sequencing, and differential expression of miRNAs was validated by quantitative real-time polymerase chain reaction analysis. The diagnostic performance of differentially expressed exosomal miRNAs for HCC was evaluated by receiver operating characteristic curve analysis. Target genes of these miRNAs were verified using luciferase reporter assays, and their functions were studied through in vitro and rescue assays. In total, 225 differentially expressed exosomal miRNAs were identified in HCC samples compared with adjacent liver tissues, and some were associated with HCC tumorigenesis and progression. Comparison of the expression profiles of tissue-derived and plasma-derived exosomal miRNAs identified hsa-miR-483-5p as the only differentially expressed miRNA detected in both HCC tissue and plasma, and this was in a validation group of HCC patients. Analysis of the diagnostic performance of plasma exosomal hsa-miR-483-5p or plasma hsa-miR-483-5p found that both could differentiate HCC and non-HCC cases. In vitro ectopic miR-483-5p expression promoted HCC cell proliferation. CDK15 was confirmed to bind with miR-483-5p directly, and thus, miR-483-5p may function by downregulating CDK15. Hsa-miR-483-5p represents a potential specific and sensitive biomarker for HCC diagnosis.

Chemotherapy-induced thrombocytopenia and platelet transfusion in patients with diffuse large B-cell lymphoma.

BACKGROUND: Currently, the risk factors associated with chemotherapy-induced thrombocytopenia (CIT) in patients with diffuse large B-cell lymphoma (DLBCL) are still undefined. Our study aimed to analyze the effects of risk factors on thrombocytopenia and to identify the threshold for infusion platelets of CIT patients who have received platelet transfusions. METHODS: We conducted a retrospective analysis of 523 patients with DLBCL from 2011 to 2013. The clinical and demographic parameters were extracted, and the risk factors associated with CIT were analyzed. The threshold for platelet transfusions in DLBCL patients with a central venous catheter (CVC) was evaluated. RESULTS: A total of 227 (43.4%) DLBCL patients had thrombocytopenia, and 63 (12.0%) had thrombocytopenia without concomitant cytopenia. We found that the choice of chemotherapy regimen was positively correlated with thrombocytopenia (P<0.001). The chemotherapy regimens most likely to result in thrombocytopenia were dexamethasone, cytarabine, cisplatin (DHAP) (92.3%), isophosphamide, carboplatin, etoposide (ICE) (89.7%), gemcitabine, dexamethasone, cisplatin (GDP) (89.7%), gemcitabine, and oxaliplatin (Gemox) (69.0%). In addition to these, high lactate dehydrogenase (LDH) (P=0.004) and Ann Arbor stage III/IV (P=0.024) were determined to be risk factors leading to thrombocytopenia. Forty patients (17.6%) had transfused platelets, and all of them were placed in the CVC. The high-threshold group (platelet count ≤20×109/L had a significantly lower amount of platelet transfusions than the low-threshold group ≤10×109/L. The platelet transfusion amount was 1.44±0.77 vs. 2.05±1.13 (P=0.047), respectively. CONCLUSIONS: The chemotherapy regimens of DHAP, ICE, GDP, and Gemox can easily lead to thrombocytopenia. A high level of LDH in the peripheral blood and Ann Arbor stage III/IV are also associated with risk factors for thrombocytopenia. A 20×109/L prophylactic platelet transfusion threshold value is safer, more effective, and thus a better choice for DLBCL patients with CVC.

Risk factors of acute myocardial infarction in middle-aged and adolescent people (< 45 years) in Yantai.

BACKGROUND: Yantai is a developed medium-sized coastal city in Eastern China, having a population of 1.6845 million. With the development of economy, some middle-aged and adolescent people (< 45 years) devote themselves to work and suffer from greater stress, which makes them ignore their own health. Moreover, they have unhealthy lifestyles and lack the knowledge of cardiovascular risk factors. OBJECTIVES: To identify the risk factors for first acute myocardial infarction in middle-aged and adolescent people in Yantai, a developed medium-sized coastal city in Eastern China. METHODS: A total of 154 consecutive patients with first acute myocardial infarction (< 45 years), were enrolled in case group, and 462 patients without myocardial infarction were enrolled in control group. Three controls with the same sex and age were matched to each case. The risk factors were identified with univariate and multivariate analysis. RESULTS: Unhealthy food habit (eating seafood and meanwhile drinking beer), hypertension, current smokers, self-perceived stress, diabetes mellitus, obesity, sleep insufficience, hypercholesterolaemia and fatigue were independent risk factors for first acute myocardial infarction (P < 0.05). CONCLUSIONS: Besides those recognized risk factors for cardiovascular disease (hypertension, hypercholesterolemia, diabetes mellitus and smoking), eating seafood and meanwhile drinking beer, self-perceived stress, sleep insufficience, obesity and fatigue were also the risk factors for first acute myocardial infarction in middle-aged and adolescent people in Yantai.