Effect of Genistein on Starch Digestion In Vitro and Its Mechanism of Action.

The digestive properties of starch are crucial in determining postprandial glycaemic excursions. Genistein, an active phytoestrogen, has the potential to influence starch digestion rates. We investigated the way genistein affected the digestive properties of starch in vitro. We performed enzyme kinetics, fluorescence spectroscopy, molecular docking, and molecular dynamics (MD) simulations for analysing the inhibitory properties of genistein on starch digestive enzymes as well as clarifying relevant mechanism of action. Our findings demonstrated that, following the addition of 10% genistein, the contents of slowly digestible and resistant starches increased by 30.34% and 7.18%, respectively. Genistein inhibited α-amylase and α-glucosidase, with half maximal inhibitory concentrations of 0.69 ± 0.06 and 0.11 ± 0.04 mg/mL, respectively. Genistein exhibits a reversible and non-competitive inhibiting effect on α-amylase, while its inhibition on α-glucosidase is a reversible mixed manner type. Fluorescence spectroscopy indicated that the presence of genistein caused declining fluorescence intensity of the two digestive enzymes. Molecular docking and MD simulations showed that genistein binds spontaneously to α-amylase via hydrogen bonds, hydrophobic interactions, and π-stacking, whereas it binds with α-glucosidase via hydrogen bonds and hydrophobic interactions. These findings suggest the potential for developing genistein as a pharmacologic agent for regulating glycaemic excursions.

Lamellar Nanoporous Metal/Intermetallic Compound Heterostructure Regulating Dendrite-Free Zinc Electrodeposition for Wide-Temperature Aqueous Zinc-Ion Battery.

Aqueous zinc-ion batteries are attractive post-lithium battery technologies for grid-scale energy storage because of their inherent safety, low cost and high theoretical capacity. However, their practical implementation in wide-temperature surroundings persistently confronts irregular zinc electrodeposits and parasitic side reactions on metal anode, which leads to poor rechargeability, low Coulombic efficiency and short lifespan. Here, this work reports lamellar nanoporous Cu/Al2Cu heterostructure electrode as a promising anode host material to regulate high-efficiency and dendrite-free zinc electrodeposition and stripping for wide-temperatures aqueous zinc-ion batteries. In this unique electrode, the interconnective Cu/Al2Cu heterostructure ligaments not only facilitate fast electron transfer but work as highly zincophilic sites for zinc nucleation and deposition by virtue of local galvanic couples while the interpenetrative lamellar channels serving as mass transport pathways. As a result, it exhibits exceptional zinc plating/stripping behaviors in aqueous hybrid electrolyte of diethylene glycol dimethyl ether and zinc trifluoromethanesulfonate at wide temperatures ranging from 25 to -30 °C, with ultralow voltage polarizations at various current densities and ultralong lifespan of >4000 h. The outstanding electrochemical properties enlist full cell of zinc-ion batteries constructed with nanoporous Cu/Al2Cu and ZnxV2O5/C to maintain high capacity and excellent stability for >5000 cycles at 25 and -30 °C.

Structure, properties, and resistant starch content of modified rice flour prepared using dual hydrothermal treatment.

In this study, modified rice flour with high resistant starch (RS) content was prepared by dual hydrothermal treatment, which combined the heat-moisture treatment with the pressure-heat treatment method. The effects of dual hydrothermal treatment on the structure and properties of modified rice flour and their relationship with RS content were further discussed. The results showed that the RS content of modified rice flour was higher than that of rice flour (RF), and dual hydrothermal treatment was more effective than single hydrothermal treatment. Adhesion and aggregation occurred between the particles of modified rice flour. Both crystallinity and short-range ordering were increased in modified rice flour compared to RF. Moreover, the modified rice flour of dual hydrothermal treatment had higher crystallinity and a more ordered short-range structure of starch, which improved RS content to a certain extent. Compared to single hydrothermal treatment, the modified rice flour of dual hydrothermal treatment had a lower viscoelasticity and a better thermal stability. Both RF and modified rice flour gels were composed mainly of free water, with minimal amounts of bound and immobile water. The study may provide a reference for the production and application of modified rice flour.

Effect of different heat-moisture treatment times on the structure, physicochemical properties and in vitro digestibility of japonica starch.

Modified starch was prepared from japonica starch (JS) by heat-moisture treatments (HMT). Under the same moisture content and HMT temperature, the effects of various HMT times on the structural, properties of JS and its in vitro digestibility properties were investigated. The results showed that adhesion occurred between the particles of japonica starch after the HMT, and there were depressions on the surface. The size of the JS particles increased, the short-range ordering and relative crystallinity of the HMT-modified starch increased and gradually decreased, reaching a peak of 36.51 % at 6 h, as the HMT time was extended. The pasting indexes of HMT-modified starch decreased and then increased with the increase of the HMT time; compared with JS, the thermal stability of HMT-modified starch increased while the pasting enthalpy decreased. All the HMT-modified starches were weakly gelatinous systems and pseudoplastic fluids. Following HMT, the amount of resistant starch (RS) and slowly digested starch (SDS) grew initially before declining. The amount of RS in HMT-modified starch peaked at 24.28 % when the HMT time was 6 h. The results of this research can serve as a theoretical foundation for the creation of modified japonica starch and its use in the food industry.

A Study on the Structural and Digestive Properties of Rice Starch-Hydrocolloid Complexes Treated with Heat-Moisture Treatment.

Rice starch-hydrophilic colloid complexes (SHCs) were prepared by incorporating xanthan gum and locust bean gum into natural rice starch. Subsequently, they underwent hygrothermal treatment (H-SHC) to investigate their structural and digestive properties with varying colloid types and added amounts of H-SHC. The results demonstrated that heat-moisture treatment (HMT) led to an increase in resistant starch (RS) content in rice starch. This effect was more pronounced after the addition of hydrophilic colloid, causing RS content to surge from 8.42 ± 0.39% to 38.36 ± 3.69%. Notably, the addition of locust bean gum had a more significant impact on enhancing RS content, and the RS content increased with the addition of hydrophilic colloids. Enzyme digestion curves indicated that H-SHC displayed a lower equilibrium concentration (C∞), hydrolysis index (HI), and gluconeogenesis index (eGI). Simultaneously, HMT reduced the solubility and swelling power of starch. However, the addition of hydrophilic colloid led to an increase in the solubility and swelling power of the samples. Scanning electron microscopy revealed that hydrophilic colloid encapsulated the starch granules, affording them protection. X-ray diffraction (XRD) showed that HMT resulted in the decreased crystallinity of the starch granules, a trend mitigated by the addition of hydrophilic colloid. Infrared (IR) results demonstrated no formation of new covalent bonds but indicated increased short-range ordering in H-SHC. Rapid viscosity analysis and differential scanning calorimetry indicated that HMT substantially decreased peak viscosity and starch breakdown, while it significantly delayed the onset, peak, and conclusion temperatures. This effect was further amplified by the addition of colloids. Rheological results indicated that H-SHC displayed lower values for G', G″, and static rheological parameters compared to natural starch. In summary, this study offers valuable insights into the development of healthy, low-GI functional foods.

Self-assembled core-shell clusters in deep eutectic solvents based on tetra-n-alkylammonium cations for high dissolution of strongly hydrogen-bonded small molecules.

Strongly hydrogen-bonded compounds such as 1,3,5-triamino-2,4,6­trinitrobenzene (TATB, an important insensitive high explosive) have excellent stability, but poor solubilities to limit their recrystallization, purification and recycling. In this study, core-shell clusters based on symmetrical tetra-n-alkylammonium [TTA]+ are designed to provide an inner cavity to incorporate TATB, and thus the clusters can separate TATB from original hydrogen-bonded networks to increase largely the solubility. Based on this design, deep eutectic solvents (DESs) based on [TTA]+ cations are first developed to yield self-assembled core-shell clusters for solubilizing TATB. Ninety-nine DESs based on [TTA]+ were prepared by combining with halide ions and hydrogen bond donors, and TATB's solubility increases with the formation of core-shell clusters. Tetrabutyl ammonium ([TBA]+ )-based DES (CS-1) displays excellent dissolution toward TATB. Room-temperature solubility of TATB in CS-1 with 32.88 mg/mL is about 10 times higher than recently reported ionic liquids and approximately 470 times higher than DMSO. Compared to traditional solvents, CS-1 shows economical and high dissolution ability toward TATB. The dissolution mechanism is demonstrated by experimental characterizations and theoretical calculations. After forming Zundel-type complexes between TATB and F-, the complexes as the core are surrounded by [TBA]+ as the shell to yield core-shell clusters through self-assembly of electrostatic interaction.

Multicenter, Randomized, Phase III Trial of Neoadjuvant Chemoradiation With Capecitabine and Irinotecan Guided by UGT1A1 Status in Patients With Locally Advanced Rectal Cancer.

PURPOSE: Differentiating the irinotecan dose on the basis of the uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1) genotype improves the pathologic complete response (pCR) rate. In this study, we further investigated preoperative irinotecan combined with capecitabine-based chemoradiotherapy for locally advanced rectal cancer. PATIENTS AND METHODS: We conducted this randomized, open-label, multicenter, phase III trial in China. Eligible patients with clinical T3-4 and/or N+ rectal adenocarcinoma, UGT1A1 genotype *1*1 or *1*28 were randomly allocated to the control group: pelvic radiation of 50 Gy/25 fractions with concurrent capecitabine, followed by oxaliplatin and capecitabine; or the experimental group: radiation with capecitabine combined with weekly irinotecan 80 mg/m2 for patients with UGT1A1*1*1 or 65 mg/m2 for patients with UGT1A1*1*28, followed by irinotecan and capecitabine. The primary end point was pCR. This trial was registered with ClinicalTrials.gov (ClinicalTrials.gov identifier: NCT02605265). RESULTS: Of the 360 patients initially enrolled, 356 were evaluated as the modified intention-to-treat population (n = 178 in both groups). Surgery was performed in 87% and 88% of patients in the control and experimental groups, respectively. The pCR rates were 15% (n = 27 of 178) and 30% (n = 53 of 178) in the control and experimental groups (risk ratio, 1.96; 95% CI, 1.30 to 2.97; P = .001). Four and 6 patients achieved complete clinical response in the control and experimental groups, respectively. Grade 3-4 toxicities were recorded in 11 (6%) and 68 (38%) patients in the control and experimental groups, respectively (P < .001). The commonest grade 3-4 toxicities were leukopenia, neutropenia, and diarrhea. The overall surgical complication rate was not significantly different between the two groups (11% v 15%; P < .001). CONCLUSION: Adding irinotecan guided by UGT1A1 genotype to capecitabine-based neoadjuvant chemoradiotherapy significantly increased complete tumor response in Chinese patients.

Contributions and Prognostic Values of N6-Methyladenosine RNA Methylation Regulators in Hepatocellular Carcinoma.

INTRODUCTION: The methylation at position N6 of adenine is called N6-methyladenosine (m6A). This transcriptional RNA modification exerts a very active and important role in RNA metabolism and in other biological processes. However, the activities of m6A associated with malignant liver hepatocellular carcinoma (LIHC) are unknown and are worthy of study. MATERIALS AND METHODS: Using the data of University of California, Santa Cruz (UCSC), the expression of M6A methylation regulators in pan-cancer was evaluated as a screening approach to identify the association of M6A gene expression and 18 cancer types, with a specific focus on LIHC. LIHC datasets of The Cancer Genome Atlas (TCGA) were used to explore the expression of M6A methylation regulators and their clinical significance. Gene Ontology (GO) analysis and Gene Set Enrichment Analysis (GSEA) were used to explore the underlying mechanism based on the evaluation of aberrant expression of m6A methylation regulators. RESULTS: The expression alterations of m6A-related genes varied across cancer types. In LIHC, we found that in univariate Cox regression analysis, up-regulated m6A modification regulators were associated with worse prognosis, except for ZC3H13. Kaplan-Meier survival curve analysis indicated that higher expression of methyltransferase-like protein 3 (METTL3) and YTH N6-methyladenosine RNA binding protein 1 (YTHDF1) genes related to the worse survival rate defined by disease-related survival (DSS), overall survival (OS), progression-free interval (PFI), and disease-free interval (DFI). Up-regulated m6A methylation regulator group (cluster2) obtained by consensus clustering was associated with poor prognosis. A six-gene prognostic signature established using the least absolute shrinkage and selection operator (LASSO) Cox regression algorithm performed better in the early (I + II; T1 + T2) stages than in the late (III + IV; T3 + T4) stages of LIHC. Using the gene signature, we constructed a risk score and found that it was an independent predictive factor for prognosis. Using GSEA, we identified processes involved in DNA damage repair and several biological processes associated with malignant tumors that were closely related to the high-risk group. CONCLUSION: In summary, our study identified several genes associated with m6A in LIHC, especially METTL3 and YTHDF1, and confirmed that a risk signature comprised of m6A-related genes was able to forecast prognosis.

lncRNA DSCAM-AS1 downregulates miR-216b to promote the migration and invasion of colorectal adenocarcinoma cells.

BACKGROUND: It has been reported that lncRNA DSCAM-AS1 plays an oncogenic role in breast cancer. In the present study we explored the role of DSCAM-AS1 in colorectal adenocarcinoma (CRA).Methods: Gene expression was analyzed by qPCR and Western blot. Overexpression experiments were performed to analyze gene interactions. Transwell assays were performed to analyze cell invasion and migration. Methylation-specific PCR (MSP) was performed to analyze DNA methylation. RESULTS: It was observed that DSCAM-AS1 was upregulated in the primary tumor tissues than in paired non-tumor tissues (within 2 cm around tumors) and was further increased with tumor metastasis. miR-216b was downregulated in primary tumor and further downregulated with tumor metastasis. miR-216b was inversely correlated with DSCAM-AS1 in tumor tissues, but not in non-tumor tissues. In cells of CRA cell lines, DSCAM-AS1 overexpression resulted in the downregulation of miR-216b, while miR-216b overexpression did not significantly affect DSCAM-AS1. DSCAM-AS1 overexpression did not significantly affect cancer cell proliferation but promoted cell migration and invasion. miR-216b inhibited cancer cell migration and invasion and significantly reduced the effects of DSCAM-AS1 overexpression. Methylation-specific PCR showed that DSCAM-AS1 overexpression promoted the methylation of miR-216b gene. CONCLUSION: DSCAM-AS1 may downregulate miR-216b to promote the migration and invasion of CRA cells.

Andrographolide suppresses proliferation of human colon cancer SW620 cells through the TLR4/NF-κB/MMP-9 signaling pathway.

Modern pharmacological research has revealed that andrographolide has various functions, including anti-bacterial, anti-inflammatory and anti-viral effects, immunoregulation, treating cardiovascular and cerebrovascular diseases, and prevention and treatment of alcoholic liver injury. The present study investigated whether andrographolide suppresses the proliferation of human colon cancer cell through the Toll-like receptor 4 (TLR4)/nuclear factor (NF)-κB/matrix metalloproteinase-9 (MMP-9) signaling pathway. The MTT assay and lactate dehydrogenase assay were used to evaluate the anticancer effects of andrographolide on cell proliferation and cytotoxicity in human colon cancer SW620 cells. Flow cytometry was used to analyze the anticancer effects of andrographolide on apoptosis by Annexin V-fluorescein isothiocyanate/propidium iodide kit. The effects of andrographolide on the activity of caspase-3/9 were measured using ELISA. Western blot analysis was also used to analyze the protein expression of TLR4, myeloid differentiation primary response gene 88 (MyD88), NF-κB-p65 and MMP-9. In the present study, it was found that andrographolide suppressed the cell proliferation, augmented cytotoxicity, evoked cell apoptosis and activated caspase-3/9 activities in human colon cancer SW620 cells. The results revealed that the anti-proliferation effects of andrographolide on the SW620 cells was associated with the inhibition of TLR4, MyD88, NF-κB-p65 and MMP-9 signaling activation. The results suggest that andrographolide is a promising drug for treatment of human colon cancer via suppression of the TLR4/NF-κB/MMP-9 signaling pathway.

miR-526b-3p functions as a tumor suppressor in colon cancer by regulating HIF-1α.

HIF-1α is an important transcriptional factor, which plays roles in cancer development and progression. But its regulation by miRNAs is not clear. Here, to investigate the regulation of HIF-1α by miRNAs, miRNAs were predicted and miR-526b-3p was verified as a regulator of HIF-1α in colon cancer cells. Using TaqMan RT-PCR analysis, we analyzed the expression of miR-526b-3p in tumor tissues and cell lines and found that miR-526b-3p was consistently under-expressed in cancer tissues and cell lines compared with their normal controls. When miR-526b-3p was induced into the colon cancer cells, cell proliferation, metastasis and glycolysis of colon cancer cells were suppressed. We also found that miR-526b-3p was down-regulated in metastatic colon cancer tissues and negatively with HIF-1α mRNA in colon cancer tissues. In a summary, miR-526b-3p plays as a tumor suppressor by down-regulation of HIF-1α expression in colon cancer and may be a new diagnosis or therapeutic target.

Fluorouracil-based preoperative chemoradiotherapy with or without oxaliplatin for stage II/III rectal cancer: a 3-year follow-up study.

BACKGROUND: Fluorouracil-based preoperative chemoradiotherapy has become the standard treatment for stage II/III rectal cancer. In order to improve the overall survival (OS) and disease-free survival (DFS), we added oxaliplatin to the standard treatment, and compared the effectiveness of these two treatment patterns. METHODS: A total of 206 patients enrolled in the prospective study had histologically confirmed rectal cancer of clinical stage II/III during July 2007 to July 2010. They were randomized into the experimental group received oxaliplatin and capecitabine in combination with radiotherapy, and the control group received capecitabine in combination with radiotherapy. All patients received surgery in 6-10 weeks after chemoradiotherapy and adjuvant chemotherapy with mFOLFOX6. The primary endpoints were DFS and OS, and the secondary endpoints included toxicity, compliance, and histopathological response. RESULTS: The 3-year OS in the experimental group and the control group was 90.29% vs. 86.41% (P>0.05), and the 3-year DFS was 80.58% vs. 69.90% (P>0.05). The pathological complete remission (pCR) rates were 23.30% and 19.42%, respectively (P=0.497). The 3-year local recurrence rates were 4.85% vs. 5.83% (P=0.694), and the 3-year distant metastasis rates were 16.50% and 28.16%, respectively (P=0.045). There were no significant differences in most grade 3-4 toxicities between two groups, however, grade 3-4 diarrhea occurred in 16.50% (17/103) of the experimental group, compared with 6.80% (7/103) of the control group (P=0.030). Also, the total grade 3-4 acute toxicity showed a significant difference (10.68% vs. 21.36%, P=0.037). CONCLUSIONS: The experimental treatment did not lead significantly improved OS and DFS, and thus longer follow-up is warranted for our patient cohort. Adding oxaliplatin to capecitabine-based preoperative chemoradiotherapy can significantly reduce metastasis, but has only minimal impact on local recurrence. Although grade 3-4 toxicity rate increased (primarily gastrointestinal toxicity), patients can stand to be followed up with allopathic treatment.

Hsp90 is involved in apoptosis of Candida albicans by regulating the calcineurin-caspase apoptotic pathway.

Candida albicans is the most common human fungal pathogen. Recent evidence has revealed the occurrence of apoptosis in C. albicans that is inducible by environmental stresses such as hydrogen peroxide, acetic acid, and amphotericin B. Apoptosis is regulated by the calcineurin-caspase pathway in C. albicans, and calcineurin is under the control of Hsp90 in echinocandin resistance. However, the role of Hsp90 in apoptosis of C. albicans remains unclear. In this study, we investigated the role of Hsp90 in apoptosis of C. albicans by using an Hsp90-compromised strain tetO-HSP90/hsp90 and found that upon apoptotic stimuli, including hydrogen peroxide, acetic acid or amphotericin B treatment, less apoptosis occurred, less ROS was produced, and more cells survived in the Hsp90-compromised strain compared with the Hsp90/Hsp90 wild-type strain. In addition, Hsp90-compromised cells were defective in up-regulating caspase-encoding gene CaMCA1 expression and activating caspase activity upon the apoptotic stimuli. Investigations on the relationship between Hsp90 and calcineurin revealed that activation of calcineurin could up-regulate apoptosis but could not further down-regulate apoptosis in Hsp90-compromised cells, indicating that calcineurin was downstream of Hsp90. Hsp90 inhibitor geldanamycin (GdA) could further decrease the apoptosis in calcineurin-pathway-defect strains, indicating that compromising Hsp90 function had a stronger effect than compromising calcineurin function on apoptosis. Collectively, this study demonstrated that compromised Hsp90 reduced apoptosis in C. albicans, partially through downregulating the calcineurin-caspase pathway.

Ascorbic acid decreases the antifungal effect of fluconazole in the treatment of candidiasis.

1. The aim of the present study was to investigate the effects of ascorbic acid (AA) on the antifungal activity of fluconazole (FCZ) in a systemic murine candidiasis model as well as in vitro. 2. The murine model was established by infusion of Candida albicans via the tail vein. Control mice received no further treatment. Other groups of mice were injected with FCZ (0.5 mg/kg, i.p.) and then treated or not with 50 or 500 mg/kg AA intragastrically (i.g.) or i.p. In all groups, FCZ was administered i.p. 2 h after fungal inoculation, whereas AA was administered 6 h after fungal inoculation. Survival rate, kidney fungal burden and renal pathological changes were evaluated. 3. The in vitro effects of AA (5, 1 and 0.2 mmol/L) on the growth of various Candida strains in the presence of FCZ (0.125-64 microg/mL) were also investigated. The in vitro effects of two anti-oxidants, namely N-acetylcysteine (NAC; 5, 1 and 0.2 mmol/L) and reduced glutathione (GSH; 5, 1 and 0.2 mmol/L), on FCZ activity were evaluated to determine the mechanism of action of AA. 4. Intragastric administration of AA (50 or 500 mg/kg) significantly decreased the antifungal effect of 0.5 mg/kg FCZ. Although i.p. administration of AA (50 or 500 mg/kg) had no significant effect on the survival of mice, it dose-dependently inhibited the activity of FCZ, with significant inhibition observed with 500 mg/kg AA. 5. In vitro, AA decreased the activity of FCZ against various Candida strains. Both NAC and GSH dose-dependently decreased the activity of FCZ. 6. The results of the present study indicate that AA inhibits the antifungal activity of FCZ, suggesting that the two should not be used together clinically for the treatment of candidiasis.

CaIPF7817 is involved in the regulation of redox homeostasis in Candida albicans.

CaIPF7817, a functionally unknown gene in Candida albicans, was suggested to be involved in the redox system previously, but its exact role is unknown. In this study, ipf7817 null mutant was generated with the URA-blaster method. After the deletion of CaIPF7817, intracellular levels of reactive oxygen species were significantly increased; mitochondrial membrane potential, a direct indicator of mitochondrial function, was elevated; some important redox-related genes, including GLR1, SOD2, and TRR1, were up-regulated; and the GSH/GSSG ratio was raised. These changes indicated that CaIPF7817 played important roles in the regulation of redox homeostasis in C. albicans.