The mechanism and experimental verification of Ixeris sonchifolia promoting apoptosis of hepatocellular carcinoma based on network pharmacology: Ixeris sonchifolia Induces Hepatocellular Carcinoma Apoptosis via the PI3K/AKT Pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Ixeris sonchifolia alias Kudiezi, it was named Ixeris sonchifolia (Bunge) Hance, a synonym for Crepidiastrum sonchifolium (Bunge) Pak & Kawano in the https://www.iplant.cn/. And it was first published in J. Linn. Soc., Bot. 13: 108 (1873), which was named Ixeris sonchifolia (Maxim.) Hance in the MPNS (http://mpns.kew.org). As a widely distributed medicinal and edible wild plant, it possesses unique bitter-cold characteristics and constituents with various pharmacological activities. Its main antitumor substances, same as artemisinin and paclitaxel, are classified as terpenoids and have become research foci in recent years. However, its specific biological activity and role in antitumor treatment remain largely unclear. AIM OF THE STUDY: This study aimed to elucidate the molecular targets and potential mechanisms of hepatocellular carcinoma apoptosis induced by Ixeris sonchifolia. MATERIALS AND METHODS: We used network pharmacology methods to analyze and screen the active ingredients and possible underlying mechanisms of Ixeris sonchifolia in treating liver cancer and employed integrative time- and dose-dependent toxicity, transcriptomics, and molecular biology approaches to comprehensively verify the function of Ixeris sonchifolia extract (IsE) in human hepatoblastoma cell (HepG2) apoptosis and its potential mechanism. RESULTS: A total of 169 common targets were screened by network pharmacology, and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that IsE inhibited HepG2 cell activity in a time- and dose-dependent manner. Western blot analysis confirmed that IsE promoted HepG2 cell apoptosis by inhibiting the PI3K/AKT signaling pathway and that the PI3K/AKT inhibitor LY294002 also substantially enhanced IsE-induced apoptosis. The PI3K/AKT signaling pathway exhibited significant differences compared to that in the control group. CONCLUSION: Combining network pharmacology with experimental verification, IsE inhibited mitochondrial function and the PI3K/AKT pathway while inducing hepatoma cell apoptosis. IsE may have promising potential for liver cancer treatment and chemoprevention.

New perspective on effect of ß-cyclodextrin on nitrification-denitrification and denitrification phosphorus removal in biogenic manganese oxides driven moving bed biofilm reactor: Performance evaluation, microbial community and process.

Effect of ß-cyclodextrin (ß-CD) on simultaneous removal of NH4+-N, NO3--N, COD, and phosphorus (P) in biogenic manganese oxides (BioMnOx) driven moving bed biofilm reactor (MBBR) was investigated. 58.64% and 86.32%, 79.65% and 98.39%, 62.45% and 97.30%, and 24.80% and 95.90% of TN and COD were removed in phases I-IV, indicating that simultaneous nitrification and denitrification (SND) efficiencies were 75.44%, 83.91%, 72.71%, and 35.83%, respectively. Composition and fluorescence spectral characteristics of extracellular polymeric substance (EPS) were evaluated including the removal kinetics of TN and COD. Metabolic activity of Mn2+, decolorization performance of BioMnOx, and reactive oxygen species (ROS) characteristics were determined in biofilm. Furthermore, intermediate Mn3+ and BioMnOx concentration were analyzed. Finally, the removal process of nitrogen (N) and P was proposed based on characterizations of elemental characterization, electrochemistry, and microbial community. This study provides new insights into the N and P removal mediated by BioMnOx and ß-CD.

Quinoxalineimide-Based Semiconducting Polymer Nanoparticles as an Effective Phototheranostic for the Second Near-Infrared Fluorescence Imaging and Photothermal Therapy.

Multifunctional theranostics play a critical role in improving the efficacy of photothermal therapy and tumor fluorescence imaging; however, they require the integration of complex components into a single theranostic system, and their response in the second near-infrared (NIR-II) region is constrained by wavelengths of a photosensitizer. To address this issue, we herein developed a novel multifunctional thiazole-fused quinoxalineimide semiconducting polymer (named PQIA-BDTT), which exhibits NIR-II fluorescence and photothermal properties. PQIA-BDTT nanoparticles achieved an impressively high photothermal conversion efficiency (72.6%) in laser (1064 nm)-induced photothermal therapy at a safe maximum permissible exposure, demonstrating their capability as an effective photothermal agent. Moreover, PQIA-BDTT nanoparticles can be used as a reference for NIR-II fluorescence imaging under a low laser fluence. The tumor size and location in 4T1 mice intravenously injected with the PQIA-BDTT nanoparticles could be precisely identified through NIR-II fluorescence imaging, which also exhibited remarkable photothermal antitumor efficacy by in vitro and in vivo therapy. Overall, this study demonstrates that introducing a thiazole-fused quinoxalineimide acceptor unit into a donor-acceptor conjugated polymer is an effective strategy for the synthesis of novel multifunctional theranostic systems, which provides a novel platform for designing theranostic agents for biomedical applications.

Overview of structure, function and integrated utilization of marine shell.

Marine shell resources have received great attention from researchers owing to their unique merits such as high hardness, good toughness, corrosion resistance, high adsorption, and bioactivity. Restricted by the level of comprehensive utilization technology, the utilization rate of shells is extremely low, resulting in serious waste and pollution. The research shows that the unique brick-mud structure of shells makes them have diverse and good functional characteristics, which guides them to have great utilization potential in different fields. Hence, this review highlights the constitutive relationship between microstructure-function-application of shells (e.g., gastropods, cephalopods, and amniotes), and the comprehensive applications and development ideas in the fields of biomedicine, adsorption enrichment, pHotocatalysis, marine carbon sink, and environmental deicer. It is worth mentioning that marine shells are currently well developed in three areas: bone repair, health care and medicinal value, and drug carrier, which together promote the progress of biomedical field. In addition, an in-depth summary of the application of marine shells in the adsorption and purification of various impurities such as crude oil, heavy metal ions and dyes at low-cost and high efficiency is presented. Finally, by integrating thoughts and approaches from different applications, we are committed to providing new pathways for the excavation and future high-value of shell resources, clarifying the existing development stages and bottlenecks, promoting the development of related technology industries, and achieving the synergistic win-win situation of economic and environmental benefits.

Overexpression of the autophagy-related gene SiATG8a from foxtail millet (Setaria italica L.) in transgenic wheat confers tolerance to phosphorus starvation.

In plants, autophagy plays an important role in regulating intracellular degradation and amino acid recycling in response to nutrient starvation, senescence, and other environmental stresses. Foxtail millet (Setaria italica) shows strong resistance to various abiotic stresses; however, current understanding of the regulation network of abiotic stress resistance in foxtail millet remains limited. In this study, we aimed to determine the autophagy-related gene SiATG8a in foxtail millet. We found that SiATG8a was mainly expressed in the stem and was induced by low-phosphorus (LP) stress. Overexpression of SiATG8a in wheat (Triticum aestivum) significantly increased the grain yield and spike number per m2 under LP treatment compared to those in the WT varieties S366 and S4056. There was no significant difference in the grain P content between SiATG8a-overexpressing wheat and WT wheat under normal phosphorus (NP) and LP treatments. However, the phosphorus (P) content in the roots, stems, and leaves of transgenic plants was significantly higher than that in WT plants under NP and LP conditions. Furthermore, the expression of P transporter genes, such as TaPHR1, TaPHR3, TaIPS1, and TaPT9, in SiATG8a-transgenic wheat was higher than that in WT under LP. Collectively, overexpression of SiATG8a increases the P content of roots, stems, and leaves of transgenic wheat under LP conditions by modulating the expression of P-related transporter gene, which may result in increased grain yield; thus, SiATG8a is a candidate gene for generating transgenic wheat with improved tolerance to LP stress in the field.

Electroacupuncture for relieving itching in atopic eczema: study protocol for a multicenter, randomized, sham-controlled trial.

Background: Atopic eczema (AE) is a common atopic inflammatory skin disease affecting 2.1-4.9% of the population in different countries. Pruritus, one of the most burdensome symptoms, is often underestimated for the problems it can cause, creating a vicious loop of itching, scratching, and lichenification. Therefore, further research into practical and safe treatments that relieve itchy symptoms and enhance skin protection is key to overcoming AE. Acupuncture, with or without electrical stimulation, is one of the most commonly used therapeutic measures to treat AE. This trial aimed to objectively evaluate the efficacy and safety of the electroacupuncture (EA) antipruritic technique in AE pruritus and obtain high-level clinical evidence for the popularization and application of EA for AE. Methods and analysis: This multicenter, single-blinded, randomized controlled trial is planned to transpire from April 15, 2023, to June 30, 2025. We will recruit 132 participants with AE (44 per group). Participants will be assigned randomly to three equal-sized groups: EA, sham electroacupuncture, and sham acupuncture. Treatment will be administered three times a week during the 2-week intervention phase. The primary outcome measure is the Visual Analog Scale, with a numeric rating scale to evaluate pruritus. Secondary outcome measures include the Eczema Area and Severity Index and Dermatology Life Quality Index. Other outcome measures include physical examination, serum IgE, and safety evaluation. The number, nature, and severity of adverse events will be carefully recorded. Trial registration: ClinicalTrials.gov, 22Y11922200. Registered 3 September 2022, https://register.clinicaltrials.gov.

Efficacy and safety of Chinese herbal medicine for atopic dermatitis: Evidence from eight high-quality randomized placebo-controlled trials.

Background: The use of Chinese herbal medicine (CHM) for the treatment of atopic dermatitis (AD) has gained attention. This quantitative study systematically evaluated the efficacy and safety of CHM for the treatment of AD in eight high-level clinical trials, resulting in a high level of clinical evidence. Methods: Several databases were searched, including PubMed, Embase, Cochrane Library, Web of Science, China National Knowledge Infrastructure (CNKI), the Chongqing VIP Chinese Science (VIP), and Wanfang Database. High-quality randomized controlled trials (RCTs) comparing CHM with placebo were included. The 95% confidence interval (CI) of the risk ratio (RR) was calculated using software (RevMan 5.3) and a meta-analysis was performed. Evidence level evaluation using GRADE Profiler 3.6. Results: In total, 662 patients (322 in the experimental group and 340 in the control group) were included. The response rate of the Eczema Area and Severity Index (EASI) -90 was higher in the CHM group than in the placebo group (RR, 3.72; 95% CI, 1.76 to7.83; p = 0.01). Furthermore, the scoring of atopic dermatitis (SCORAD) (RR, -10.20), body surface area (BSA) (RR, -2.01), surface damage score (RR, -2.25), visual analog scale (VAS) (RR, -1.90), and sleep score (RR, -2.16), improvement of investigator's global assessment (IGA) (RR, 2.94) improved in the CHM group. The results showed no statistical difference between CHM and placebo (MD, -0.47; 95% CI, -1.30, 0.37; p = 0.27) in improving the Dermatology Life Quality Index (DLQI) or children's DLQI (CDLQI). There was also no significant difference in the IgE level between the two groups (MD, -62.76; 95% CI, -809.58, 684.05; p = 0.87). However, the adverse events (AEs) rate was slightly higher in patients treated with CHM than in those treated with placebo (RR, 1.42; 95% CI, 1.06-1.90; p = 0.02). Conclusion: CHM improved the size and severity of the skin lesions and sleep quality in patients with AD. Comparing the adverse effects between the two groups, CHM is safe. However, CHM does not improve the quality of life or the patient's IgE levels.

Ramucirumab for Patients with Advanced Hepatocellular Carcinoma and Elevated Alpha Fetoprotein Following Non-Sorafenib Systemic Therapy: An Expansion Cohort of REACH-2.

BACKGROUND: Ramucirumab is indicated for patients with advanced hepatocellular carcinoma (HCC) and α-fetoprotein (AFP) ≥400 ng/mL following sorafenib. Here, we prospectively studied ramucirumab following non-sorafenib systemic therapies. MATERIALS AND METHODS: This open-label, non-comparative cohort of REACH-2 enrolled patients with advanced HCC, Child-Pugh class-A liver disease, and AFP ≥400 ng/mL who had received 1-2 lines of therapy, excluding sorafenib or chemotherapy. Ramucirumab was administered 8 mg/kg intravenously Q2W. The primary endpoint was safety. Secondary endpoints were overall survival, progression-free survival, objective response rate (RECIST v1.1), time to progression, pharmacokinetics, and patient-reported outcomes. Final analysis occurred after all enrolled patients completed ≥3 treatment cycles or discontinued treatment. RESULTS: Between April 27, 2018, and March 29, 2021, 47 patients were treated at 21 investigative sites in Asia, Europe, and USA. The most frequently reported grade ≥3 adverse events, regardless of causality, were hypertension (11%), proteinuria (6%), hyponatremia (6%), and AST increased (6%). Two patients died from adverse events (myocardial infarction and upper gastrointestinal hemorrhage), deemed related to treatment. Median progression-free survival, time to progression, and overall survival were 1.7 months, 2.8 months, and 8.7 months, respectively. The objective response rate was 10.6% with a median duration response of 8.3 months. Median time to deterioration in FHSI-8 total score was 4.4 months. CONCLUSION: Ramucirumab demonstrated consistent and meaningful clinical activity with no new safety signals following non-sorafenib therapies in patients with advanced HCC and AFP ≥400 ng/mL. This represents one of the first sequencing studies for patients with advanced HCC not treated with sorafenib.

Cutaneous allergic reaction to subcutaneous vitamin K1: A case report and review of literature.

BACKGROUND: Vitamin K1 (phytomenadione) is a fat-soluble naturally occurring vitamin that is widely used to treat certain coagulation disorders. Adverse cutaneous reactions to vitamin K1 can occur; however, owing to its low incidence and considerable variability in presentation and morphology, its diagnosis can be easily overlooked. Managing these reactions may be challenging for patients and clinicians. Therefore, reviewing the adverse cutaneous reactions to vitamin K1 is important. CASE SUMMARY: Here we report the case of a 50-year-old woman with no pre-existing hepatic disease who developed a cutaneous allergic reaction to subcutaneous vitamin K1 that presented as localized eczematous plaques at the vitamin K1 injection site. The eruption developed within 5 d of the injection and persisted for 32 mo despite treatment with topical and intralesional steroids. Eczema was diagnosed based on the results of the pathological examination, immunohistochemical staining, and a skin biopsy. The patient was advised to take herbal medicines orally twice daily. After treatment and follow-up, the patient's eczematous urticarial plaques improved and her condition stabilized. CONCLUSION: Here we present the first case of a cutaneous allergic reaction to subcutaneous vitamin K1 that was successfully treated with Chinese medicine.

Performance of simultaneous nitrification-denitrification and denitrifying phosphorus and manganese removal by driving a single-stage moving bed biofilm reactor based on manganese redox cycling.

Simultaneous removal of NH4+-N, NO3--N, COD, and P by manganese redox cycling in nutrient wastewater was established with a single-stage moving bed biofilm reactor (MBBR) under low C/N ratio. When sodium succinate replaced the conventional denitrifying carbon source, removal efficiencies of TN, NO3--N, NH4+-N, TP, and Mn2+ were 65.13 %, 79.63 %, 92.79 %, 51.57 %, and 68.10 %, respectively. Based on modified Stover-Kincannon model, 11.03 and 10.05 mg TN·L-1·h-1 of Umax values were obtained with sodium acetate and sodium succinate as substrates. Extracellular polymeric substances were used to evaluate the characteristics of biofilm, and microbial community of biofilm was identified. Transformation processes of NO3--N, NH4+-N, Mn2+, and P were investigated, suggesting that the main functional groups (e.g., CO, Mn-O, and CN bonds) participated in N, P, and Mn2+ removal, and MnO2 was the main component of biogenic manganese oxides. This study provides a new strategy for nutrients removal by Mn2+ driven MBBR.

Treatment heterogeneity and overall survival in patients with advanced/metastatic gastric or gastroesophageal junction adenocarcinoma in the United States.

Background: Gastric or gastroesophageal junction (GEJ) adenocarcinoma is the most common form of gastric cancer diagnosed in the United States (US) each year. Diagnosis typically is in later stages of disease when it has advanced. Patients have been treated with a variety of regimens. Methods: The goal of this retrospective study was to understand if treatment patterns were becoming more homogeneous or remaining heterogeneous using the Herfindahl-Hirschman index (HHI) and if treatments were becoming more concordant to treatment guidelines published by the National Comprehensive Cancer Network (NCCN). HHI scores were calculated for each site by 2-year increments. Trend analyses were conducted for HHI scores over time using a linear regression model. Concordance to Category 1 and any category NCCN guidelines was determined based on the date treatment was initiated with the version of the NCCN guidelines at that time. Time trend analyses were conducted using linear regression models. This study utilized data from the Flatiron Advanced Gastric/Esophageal cohort. This study also examined overall survival (OS) rates estimated by the Kaplan-Meier method by line of therapy. Results: There were no statistically significant differences in HHI scores in the first-line setting over time, suggesting heterogeneity has not improved. Concordance to NCCN treatment guidelines for any category significantly increased over time, however Category 1 regimen concordance remained low in the first-line setting. Concordance over time improved in second-line treatment. Median OS from the start of first-line therapy was 13.57 months. There was no relationship between OS time from initiation of first-line therapy and HHI score, concordance with NCCN guidelines, or concordance with NCCN Category 1 guidelines in the first-line setting. Conclusions: Treatment heterogeneity persists in gastric cancer care, though there is a significant association between heterogeneity and concordance with both Category 1 and any category in the NCCN treatment guidelines, and that concordance has increased over time.

Isoorientin ameliorates OVA-induced asthma in a murine model of asthma.

Allergic asthma which is induced by ovalbumin (OVA) is a chronic airway inflammation disease. Isoorientin (Iso) is a natural C-glucosyl flavone with many biological properties. We aimed to evaluate the effectiveness of Iso on OVA-induced allergic asthma. A total of 30 C57BL/6 mice were randomly divided into five groups: control group, OVA group, Dex (dexamethasone, 10 mg/kg) group, low-dose Iso group (Iso-L, 25 mg/kg), and high-dose Iso group (Iso-H, 50 mg/kg). The serum and bronchoalveolar lavage fluid (BALF) were collected for biochemical parameters, the lung tissue was collected for hematoxylin-eosin (H&E) staining, immunohistochemistry (IHC), and western blot. The levels of IL-4, IL-5, IL-13, malondialdehyde (MDA), NO, and reactive oxygen species (ROS) in Iso-L and Iso-H groups were significantly lower than that in model group (p < 0.05). Simultaneously, the levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity were higher than that in model group (p < 0.05). Iso significantly ameliorated airway hyperresponsiveness. Meanwhile, H&E staining revealed that mice treated with Iso resulted in the ameliorated inflammatory cell infiltration and a reduction in interstitial thickening. The nuclear factor erythroid 2-like 2 (Nrf2) and HO-1 protein expression in Iso-L and Iso-H groups were enhanced over that in model group, while p-NF-κB-p65 and p-IκB-α protein expression was decreased (p < 0.05). Our research indicated that Iso alleviated the OVA-induced allergic asthma, and this effect can be explained by the modulation of Nrf2/HO-1 and NF-κB signaling pathway; thus, the results providing a therapeutic rationale for the treatment of Iso on allergic asthma.

Auriculasin enhances ROS generation to regulate colorectal cancer cell apoptosis, ferroptosis, oxeiptosis, invasion and colony formation.

Colorectal cancer (CRC) is one of the most common malignant tumors in the digestive system, and Chinese herbal medicine plays an important role in tumor treatment. The in-depth study of auriculasin isolated from Flemingia philippinensis showed that auriculasin promoted reactive oxygen species (ROS) generation in a concentration-dependent manner; when ROS scavenger NAC was added, the effects of auriculasin in promoting ROS generation and inhibiting cell viability were blocked. Auriculasin induced CRC cell apoptosis, led to mitochondrial shrinkage, and increased the intracellular accumulation of Fe2+ and MDA. When auriculasin and NAC were added simultaneously, the levels of apoptosis, Fe2+ and MDA returned to the control group levels, indicating that auriculasin activated apoptosis and ferroptosis by inducing ROS generation. In addition, auriculasin promoted the expression of Keap1 and AIFM1, but significantly reduced the phosphorylation level of AIFM1, while NAC significantly blocked the regulation of Keap1 and AIFM1 by auriculasin, which indicates that auriculasin can also induce oxeiptosis through ROS. When Z-VAD-FMK, Ferrostatin-1, Keap1 siRNA, PGAM5 siRNA and AIFM1 siRNA were added respectively, the inhibitory effect of auriculasin on cell viability was significantly weakened, indicating that auriculasin inhibits cell viability by inducing apoptosis, ferroptosis and oxeiptosis. Auriculasin also inhibited the invasion and clone forming ability of CRC cells, while NAC blocked the above effects of auriculasin. Therefore, auriculasin can promote CRC cell apoptosis, ferroptosis and oxeiptosis by inducing ROS generation, thereby inhibiting cell viability, invasion and clone formation, indicating that auriculasin has a significant antitumor effect.

Exploration of and insights into advanced topical nanocarrier systems for the treatment of psoriasis.

Psoriasis is a chronic inflammatory skin disease with an underlying autoimmune pathogenesis that has brought great distress to patients. Current treatment options include topical therapy, systemic therapy, and phototherapy. By disrupting the stratum corneum, nanocarriers have unique advantages in allowing drug carriers to be tailored to achieve targeted drug delivery, improve efficacy, and minimize adverse effects. Furthermore, despite their limited success in market translatability, nanocarriers have been extensively studied for psoriasis, owing to their excellent preclinical results. As topical formulations are the first line of treatment, utilize the safest route, and facilitate a targeted approach, this study, we specifically describes the management of psoriasis using topical agents in conjunction with novel drug delivery systems. The characteristics, advantages, weaknesses, and mechanisms of individual nanocarriers, when applied as topical anti-psoriatic agents, were reviewed to distinguish each nanocarrier.

Aggregation-induced emission (AIE)-guided dynamic assembly for disease imaging and therapy.

The phenomenon of aggregation-induced emission (AIE) is inseparable from molecular aggregation and self-assembly. Therefore, the combination of AIE and supramolecular self-assembly is well-matched. AIE-guided dynamic assembly (AGDA) could effectively respond to the endogenous stimuli (such as pH, enzymes, redox molecules) and exogenous stimuli (temperature, light, ultrasound) in the disease microenvironment, so as to achieve specific imaging and diagnosis of the disease lesions. Moreover, AGDA also dynamically adjust the intramolecular motions of AIE molecules, thereby adjusting the energy dissipation pathways and realizing the switch between photodynamic therapy and photothermal therapy for superior therapeutic effects. In this review, we aim to give an overview of the constructing strategies, stimuli-responsive imaging, regulation of intramolecular motion of AGDA in recent years, which is expected to grasp the research status and striving directions of AGDA for imaging and therapy.

Protosappanin B Exerts Anti-tumor Effects on Colon Cancer Cells via Inhibiting GOLPH3 Expression.

Protosappanin B (PSB) is a key active component of Lignum Sappan extract. Although the antiproliferative effects of Lignum Sappan extract have been demonstrated in various cancer cells, relatively little is known about the effects of PSB on tumor progression. The aim of this study was to explore the anti-tumor effects of PSB on human colon cancer cells by regulation of intracellular signaling pathways and Golgi phosphoprotein 3 (GOLPH3) expression in vitro and in vivo. Our results showed that PSB effectively inhibited the viability and migration of SW620 cells and induced apoptosis, but had poor effect on HCT116 cells. Furthermore, PSB significantly reduced the expression of p-AKT, p-p70S6K, ß-catenin, and p-ERK1/2 proteins in SW620 cells, and this effect was reversed by the corresponding signaling pathway agonists. Interestingly, PSB could also suppress GOLPH3 expression of SW620 cells in a concentration-dependent manner, but SW620 cells transfected with lentiviral vectors overexpressing GOLPH3 can effectively resist the cytotoxic activity of PSB in vitro. The xenograft experiment of SW620 cells with LV-GOLPH3 confirmed that PSB distinctly inhibited the tumor growth via suppressing GOLPH3 expression. Collectively, these findings clarified a new anti-cancer mechanism of PSB through inhibition of GOLPH3 expression and intracellular signaling pathways in colon cancer cells. PSB may be a potential new drug for colon cancer.

Tetramethylpyrazine from adlay (Coix lacryma-jobi) biotransformation by Bacillus subtilis and its quality characteristics.

Adlay, as a traditional Chinese medicine, has been used in nourishing foods, which are rich in a variety of nutrients (special biological compounds). The study was designed to optimize the fermentation parameters of dehulled, polished and broken adlay fermented by Bacillus subtilis BJ3-2 with regard to tetramethylpyrazine (TMP) yield and fibrinolytic enzyme activity. Then the proximate and bioactive components of B. subtilis-fermented adlay were evaluated. Box-Behnken design results showed that the TMP yield was 6.93 mg/g DW (dried weight) of B. subtilis-fermented polished adlay, which was about 136 times higher than that of B. subtilis-fermented soybean (BSB). The fibrinolytic enzyme activity was 2236.17 U/g in B. subtilis-fermented dehulled adlay, and slightly less than in BSB. B. subtilis-fermented adlay contained higher fat, free amino acids and fatty acids contents but lower protein and starch contents than raw adlay. Except for coixol and coixan, the levels of γ-aminobutyric acid, triterpenes, phenolics, flavonoids and coixenolide in B. subtilis-fermented adlay increased by 14.05, 2.02, 2.31 and 1.36 times, respectively. The contents of phenolic acids including caffeic, gallic, catechinic and chlonogenic acids in the free phenolic extracts significantly increased (p < 0.05). The results demonstrated that the biotransformation of high-yield TMP, fibrinolytic enzyme and other bioactive components of B. subtilis-fermented adlay products was realized. B. subtilis-fermented adlay could be a promising value-added food, and that is more suitable for human consumption.

Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene.

OBJECTIVE: Tenacissoside H (TDH) is a Chinese medicine monomer extracted from Marsdenia tenacissima extract (MTE), which has been confirmed to have antitumor effects, but its mechanism is still unclear. The aim of this study was to investigate the effect and mechanism of TDH on human colon cancer LoVo cell proliferation and migration and explore the correlation of TDH treatment with the expression of GOLPH3 and cell signaling pathways in LoVo cells. METHODS: LoVo cells were treated with TDH at 0.1, 1, 10, and 100 µg/mL for 24, 48, and 72 h. The proliferation rate of LoVo cells was evaluated by MTT assay. Recombinant plasmid p-CMV-2-GOLPH3 was constructed, and p-CMV-2-GOLPH3 and p-CMV-2 empty plasmids were transfected into LoVo cells by lipofection. Western blotting was used to detect the transfection efficiency and the expression of p-p70S6K, p70S6K, ß-catenin, and GOLPH3. The apoptosis rate was analyzed with Annexin V-FITC/PI double-staining method, and cell migration assessed by transwell assay. RESULTS: TDH inhibited the proliferation of LoVo cells in a concentration-dependent manner. The IC50 of TDH treatment in LoVo cells at 24, 48, and 72 h was 40.24, 13.00, and 5.73 µg/mL, respectively. TDH treatment significantly induced apoptosis and suppressed the viability and migration of human colon cancer LoVo cells. The effect of TDH on induction of apoptosis and inhibition of migration in LoVo cells decreased significantly after activating the PI3K/AKT/mTOR and Wnt/ß-catenin signaling pathways with agonists. Additionally, the expression of GOLPH3 protein downregulated significantly in LoVo cells under TDH treatment. Overexpression of the GOLPH3 gene increased the expression of key proteins in PI3K/AKT/mTOR and Wnt/ß-catenin signaling pathways and blocked the antitumor activity of TDH. CONCLUSION: Collectively, the present results indicated that TDH can inhibit the proliferation vitality of colon cancer LoVo cells through downregulating GOLPH3 expression and activity of PI3K/AKT/mTOR and Wnt/ß-catenin signaling pathways.

Anti-tumor Effect of Rhaponticum uniflorum Ethyl Acetate Extract by Regulation of Peroxiredoxin1 and Epithelial-to-Mesenchymal Transition in Oral Cancer.

Objective: To explore whether Rhaponticum uniflorum (R. uniflorum) had anti-tumor effects in oral cancer and investigate the molecular mechanisms involved in these anti-tumor effects. Methods: Chemical compositions of R. uniflorum ethyl acetate (RUEA) extracts were detected by ultra-performance liquid chromatography-Q/time-of-flight mass spectrometry (UPLC-Q/TOF-MS), followed by pharmacology-based network prediction analysis. The effects of RUEA extracts on proliferation, apoptosis, migration, and invasion ability of human oral squamous cell carcinoma (OSCC) cell line SCC15 were evaluated by CCK8 assay, Annexin V- fluorescein isothiocyanate/propidium iodide staining, wound healing assay, and Matrigel invasion assay, respectively. The mRNA and protein expression of peroxiredoxin1 (Prx1), the epithelial-to-mesenchymal transition (EMT) marker E-cadherin, vimentin, and Snail were determined by quantitative real-time reverse transcription polymerase chain reaction and western blotting. A mouse xenograft model of SCC15 cells was established to further evaluate the effect of RUEA extracts in vivo. Immunohistochemical assessment of Ki67 and terminal deoxynucleotidyl transferase dUTP nick end labeling staining of apoptotic cells were performed on the tumor tissues to assess the effects of RUEA extracts on proliferation and apoptosis. Results: Fourteen compounds were identified from RUEA extracts by UPLC-Q/TOF-MS. The pharmacology-based network prediction analysis showed that Prx1 could be a potential binder of RUEA extracts. In SCC15 cells, RUEA extracts inhibited cell viability, induced apoptosis, and suppressed cell invasion and migration in a concentration-dependent manner. After treatment with RUEA extracts, the mRNA and protein expression of E-cadherin increased, whereas those of Prx1, vimentin, and Snail decreased. RUEA extracts also affected the EMT program and suppressed cell invasion and migration in Prx1 knockdown SCC15 cells. In an OSCC mouse xenograft model, RUEA extracts (25 and 250 mg/kg) significantly inhibited the growth of tumors. Compared with the control group, Ki67 expression was reduced and apoptosis rates were elevated in the transplanted tumors treated with RUEA extracts. RUEA extracts increased the expression of E-cadherin and decreased the expression of Prx1, vimentin, and Snail in vivo. Conclusion: RUEA extracts inhibited tumor growth and invasion by reducing Prx1 expression and suppressing the EMT process in OSCC. RUEA extracts may be a potential candidate for OSCC treatment.