Structural characterization and binding interaction of rice glutelin fibrils complexing with curcumin.

The rice glutelin (RG), the separated retentate (RGFs) and filtrate (FGFs) fractions from total glutelin fibrils (TGFs) at pH 3.5 were used as carrier for curcumin in this test. The solubility and antioxidant activities of curcumin were improved after binding with protein and fibrils. Compared to other complexes, the RGFs-curcumin complex exhibited a highest curcumin solubility (48.05%) and a superior sustained release property, probably owing to the stable hydrogen bond between the surface groups of fibrils and hydroxyl groups of polyphenols. In addition, thermodynamic parameters revealed that the RG/TGFs/RGFs-curcumin complexes were stabilized by hydrogen bonds and van der Waals forces, whereas FGFs interacted with curcumin through specific electrostatic interaction. Besides, after interacting with curcumin, the fibrils gathered into coarsened and agglutinated fibrillar aggregates, relating to the increment of a-helix and ß-sheet structure. These results suggested that RGFs could be a good alternative for curcumin delivery in food industry.

EGFR/MAPK signaling pathway acts as a potential therapeutic target for sulforaphane-rescued heart tube malformation induced by various concentrations of PhIP exposure.

BACKGROUND: 2-Amino-1-methyl-6-phenylimidazo [4,5-b] pyrimidine (PhIP) is a known carcinogen generated mainly from cooking meat and environmental pollutants. It is worth exploring the potential of natural small-molecule drugs to protect against adverse effects on embryonic development. PURPOSE: In this study, we investigated the potential toxicological effects of PhIP on embryonic heart tube formation and the effect of Sulforaphane (SFN) administration on the anti-toxicological effects of PhIP on embryonic cardiogenesis. STUDY DESIGN AND METHODS: First, the chicken embryo model was used to investigate the different phenotypes of embryonic heart tubes induced by various concentrations of PhIP exposure. We also proved that SFN rescues PhIP-induced embryonic heart tube malformation. Second, immunofluorescence, western blot, Polymerase Chain Reaction (PCR) and flow cytometry experiments were employed to explore the mechanisms by which SFN protects cardiac cells from oxidative damage in the presence of PhIP. We used RNA-seq analysis, molecular docking, in situ hybridization, cellular thermal shift assay and solution nuclear magnetic resonance spectroscopy to explore whether SFN protects cardiogenesis through the EGFR/MAPK signaling pathway. RESULTS: The study showed that PhIP might dose-dependently interfere with the C-looping heart tube (mild) or the fusion of a pair of bilateral endocardial tubes (severe) in chick embryos, while SFN administration prevented cardiac cells from oxidative damage in the presence of high-level PhIP. Furthermore, we found that excessive reactive oxygen species (ROS) production and subsequent apoptosis were not the principal mechanisms by which low-level PhIP induced malformation of heart tubes. This is due to PhIP-disturbed Mitogen-activated protein kinase (MAPK) signaling pathway could be corrected by SFN administration. CONCLUSIONS: This study provided novel insight that PhIP exposure could increase the risk of abnormalities in early cardiogenesis and that SFN could partially rescue various concentrations of PhIP-induced abnormal heart tube formation by targeting EGFR and mediating EGFR/MAPK signaling pathways.

Removal of typical pollutant ciprofloxacin using iron-nitrogen co-doped modified corncob in the presence of hydrogen peroxide.

Iron-nitrogen co-doped modified corncob (Fe-N-BC) was synthesized using a hydrothermal and calcination method. The material shows excellent oxidation performance and environmental friendliness. When the dosage of Fe-N-BC was 0.6 g L-1, the concentration of H2O2 was 12 mM and pH was 4, ciprofloxacin (CIP) was virtually totally eliminated in 240 min under Fe-N-BC/H2O2 conditions. The TOC removal efficiency was 54.6%, and the effects of various reaction parameters on the catalytic activity of Fe-N-BC were thoroughly assessed. Through electron paramagnetic resonance (EPR) analyses and free radical quenching experiments, it was established that the reactive oxygen species (˙OH, ˙O2-, 1O2) were crucial in the elimination of CIP. Furthermore, the degradation of CIP was accelerated by the synergistic interaction between the transition metal and PFRs. A thorough evaluation was conducted to assess the respective contributions of adsorption and catalytic oxidation in the system. The degradation mechanism of CIP was proposed under Fe-N-BC/H2O2 conditions. Meanwhile, the possible degradation intermediates and pathways were proposed, and the toxicity of the degradation products of CIP was also meticulously investigated in the study. These findings offered the elimination of CIP in water a theoretical foundation and technical support.

CD40-targeting magnetic nanoparticles for MRI/optical dual-modality molecular imaging of vulnerable atherosclerotic plaques.

BACKGROUND AND AIMS: Acute coronary syndrome caused by vulnerable plaque rupture or erosion is a leading cause of death worldwide. CD40 has been reported to be highly expressed in atherosclerotic plaques and closely related to plaque stability. Therefore, CD40 is expected to be a potential target for the molecular imaging of vulnerable plaques in atherosclerosis. We aimed to design a CD40-targeted magnetic resonance imaging (MRI)/optical multimodal molecular imaging probe and explore its ability to detect and target vulnerable atherosclerotic plaques. METHODS: CD40-Cy5.5 superparamagnetic iron oxide nanoparticles (CD40-Cy5.5-SPIONs), which comprise a CD40-targeting multimodal imaging contrast agent, were constructed by conjugating CD40 antibody and Cy5.5-N-hydroxysuccinimide ester with SPIONs. During this in vitro study, we observed the binding ability of CD40-Cy5.5-SPIONs with RAW 264.7 cells and mouse aortic vascular smooth muscle cells (MOVAS) after different treatments, using confocal fluorescence microscopy and Prussian blue staining. An in vivo study involving ApoE-/- mice fed a high-fat diet for 24-28 weeks was performed. 24 h after intravenous injection of CD40-Cy5.5-SPIONs, fluorescence imaging and MRI were performed. RESULTS: CD40-Cy5.5-SPIONs bind specifically to tumor necrosis factor (TNF)-α-treated macrophages and smooth muscle cells. Fluorescence imaging results showed that, compared with the control group and the atherosclerosis group injected with non-specific bovine serum albumin (BSA)-Cy5.5-SPIONs, the atherosclerotic group injected with CD40-Cy5.5-SPIONs had a stronger fluorescence signal. T2-weighted images showed that the carotid arteries of atherosclerotic mice injected with CD40-Cy5.5-SPIONs had a significant substantial T2 contrast enhancement effect. CONCLUSIONS: CD40-Cy5.5-SPIONs could potentially serve as an effective MRI/optical probe for vulnerable atherosclerotic plaques during non-invasive detection.

N6-Methyladenosine Demethylase FTO (Fat Mass and Obesity-Associated Protein) as a Novel Mediator of Statin Effects in Human Endothelial Cells.

BACKGROUND: N6-methyladenosine (m6A) plays a critical role in various biological processes. However, no study has addressed the role of m6A modification in the statin-induced protection of endothelial cells (ECs). METHODS: Quantitative real-time polymerase chain reaction and Western blotting analyses were used to study the expression of m6A regulatory genes in atorvastatin-treated ECs. Gain- and loss-of-function assays, methylated RNA immunoprecipitation analysis, and dual-luciferase reporter assays were performed to clarify the function of FTO (fat mass and obesity-associated protein) in ECs. RESULTS: Atorvastatin decreased FTO protein expression in ECs. The knockdown of FTO enhanced the mRNA and protein expression of KLF2 (Kruppel-like factor 2) and eNOS (endothelial NO synthase) but attenuated TNFα (tumor necrosis factor alpha)-induced VCAM-1 (vascular cell adhesion molecule 1) and ICAM-1 (intercellular adhesion molecule 1) expression, as well as the adhesion of monocytes to ECs. Conversely, FTO overexpression significantly upregulated the mRNA and protein levels of VCAM-1 and ICAM-1, downregulated those of KLF2 and eNOS, and strongly attenuated the atorvastatin-mediated induction of KLF2 and eNOS expression. Subsequent investigations demonstrated that KLF2 and eNOS are functionally critical targets of FTO. Mechanistically, FTO interacted with KLF2 and eNOS transcripts and regulated their expression in an m6A-dependent manner. After FTO silencing, KLF2 and eNOS transcripts with higher levels of m6A modification in their 3' untranslated regions were captured by YTHDF3 (YT521-B homology m6A RNA-binding protein 3), resulting in mRNA stabilization and the induction of KLF2 and eNOS protein expression. CONCLUSIONS: FTO might serve as a novel molecular target to modulate endothelial function in vascular diseases.

Complexation of rice glutelin fibrils with cyanidin-3-O-glucoside at acidic condition: Thermal stability, binding mechanism and structural characterization.

The complexation of rice glutelin fibrils (RGFs) with cyanidin-3-O-glucoside (C3G) at acidic condition was investigated. The RGFs at pH 3.5 had a greatest protective effect on the thermal stability of C3G. The binding of C3G for RGFs was exothermic and driven by hydrophobic and electrostatic interactions. The RGFs exhibited a stronger binding interaction with C3G than rice glutelin (RG), resulting from the exposure of hydrophobic groups and positive charges on the fibrils surface, and thus RGFs exhibited better protective effect on C3G. The interaction with C3G resulted in the rearrangement of polypeptide chain, thereby reducing the ß-sheet content. The larger aggregates were observed in RG/RGFs-C3G complexes due to protein-polyphenols aggregation. It was noteworthy that the pre-formed RGFs were restructured into entangled aggregates due to the interaction. This study proposed a novel protein fibril to protect anthocyanins, expanding the application of anthocyanins as stable and functional ingredients in acidic food systems.

Meta-analysis-based systematic review of effect of traditional Chinese medicine intervention in treatment of diabetic nephropathy on thyroid function.

BACKGROUND: This research sought to systematically evaluate the clinical effects of traditional Chinese medicine (TCM) intervention in the treatment of diabetic nephropathy (DN) and analyze changes in thyroid function in patients with DN. METHODS: The PubMed, Embase, Medline, Ovid, Springer, and self-built databases were searched to screen literature on TCM intervention and the treatment of DN published from the establishment of the databases to January 1, 2021. The Cochrane Handbook for Systematic Reviews of Intervention 5.0.2 was then employed to assess the risk of bias in literature, and Review Manager 5.3 was utilized for the meta-analysis. RESULTS: A total of 20 randomized controlled trials (RCTs) were included in the study, involving 3,566 subjects, and meta-analysis results showed that the clinical treatment efficiency of the experimental group was dramatically higher than the control group [MD =6.22, 95% confidence interval (CI): 3.77-10.25, Z=7.17, P<0.00001]. Moreover, the serum creatinine (Scr), blood urea nitrogen (BUN), urine protein excretion rate (UAER), 24 h postoperative urine protein quantification, and tumor necrosis factor-alpha (TNF-α) of patients after TCM intervention were all remarkably inferior to those of the control group as seen in the following results: Scr, MD =-8.69, 95% CI: -9.92 to -7.47, Z=13.94, P<0.00001; BUN, MD =-1.74, 95% CI: - 2.48 to -1.00, Z=4.6, P<0.00001; UAER, MD =-26.16, 95% CI: -46.89 to -5.44, Z=2.47, P=0.01; 24 h postoperative urine protein quantification, MD =-0.54, 95% CI: -0.68 to -0.4, Z=7.4, P<0.00001; TNF-α, MD =-5.3, 95% CI: -9.15 to -1.46, Z=2.7, P=0.007; and high sensitivity C-reactive protein (hs-CRP), MD =-1.34, 95% CI: -1.9 to -0.78, Z=4.66, P<0.00001. DISCUSSION: TCM intervention in DN is effective in treating the clinical symptoms of patients with this disease and has ideal therapeutic effects.

Rice peptide nanoparticle as a bifunctional food-grade Pickering stabilizer prepared by ultrasonication: Structural characteristics, antioxidant activity, and emulsifying properties.

In this study, a novel food-grade Pickering stabilizer was fabricated from insoluble rice peptide aggregates that are considered undesirable and formed during the hydrolysis of rice protein using ultrasonication. The results confirmed that ultrasonication was effective in fabricating rice peptide nanoparticles (RPNs) with a spherical appearance, and the particle size was reduced with ultrasonic time, reaching a minimum size of 357.8 nm in 30 min. Moreover, ultrasonic treatment could improve the antioxidant activity of RPNs by promoting the DPPH scavenging (3.5-fold increase) and Fe2+ chelating activity (3.8-fold increase). Notably, the bioactive RPNs could form stable Pickering emulsions that possess both physical and oxidative stability during storage, which might be due to the antioxidative physical barrier formed by RPNs. These findings suggest a new approach for the effective utilization of insoluble aggregates produced during protein hydrolysis as well as provide a novel bifunctional Pickering stabilizer with intrinsic antioxidant properties.

Acupuncture for thyroid nodule treatment: A protocol of systematic review and meta-analysis of randomized clinical trials.

INTRODUCTION: Thyroid nodules are scattered lesions caused by abnormal local growth of thyroid cells. In recent years, their prevalence rate has been rising gradually, and the probability of cancerations has also been increasing gradually. Therefore, we must pay more attention to them and carry out early intervention. However, at present, most of the intervention measures for patients with thyroid nodules are mainly clinical observation and follow-up, and no clear and effective drug intervention therapy has been proposed. The curative effect of acupuncture on thyroid nodules has been proved clinically. However, as there is no clear mechanism of action, no specific operation methods or Suggestions, it is necessary to make a systematic evaluation of acupuncture therapy, so as to lay a foundation for further research in the future. METHODS AND ANALYSIS: The following databases will be searched from their inception to June 2020: Electronic database includes PubMed, Embase, Cochrane Library, Web of Science, Nature, Science online, Chinese Biomedical Database WanFang, VIP medicine information, and China National Knowledge Infrastructure (CNKI). Primary outcomes: Color ultrasound of thyroid and cervical lymph nodes, FT3, FT4, TSH, TGAB, TPOAB, insulin resistance index (HOMA-IR). Data will be extracted by 2 researchers independently, risk of bias of the meta-analysis will be evaluated based on the Cochrane Handbook for Systematic Reviews of Interventions. All data analysis will be conducted by data statistics software Review Manager V.5.3. and Stata V.12.0. RESULTS: The results of this study will systematically evaluate the efficacy and safety of acupuncture therapy for patients with thyroid nodule. CONCLUSION: Through the systematic review of this study, the evidence of the treatment of thyroid nodule by acupuncture has been summarized so far, so as to provide guidance for further promoting the application of acupuncture therapy in patients with thyroid nodule. ETHICS AND DISSEMINATION: This study is a systematic review, the outcomes are based on the published evidence, so examination and agreement by the ethics committee are not required in this study. We intend to publish the study results in a journal or conference presentations. OPEN SCIENCE FRA NETWORK (OSF) REGISTRATION NUMBER: August 18, 2020. osf.io/uzck4. (https://osf.io/uzck4).

Preservation of hydrogen peroxide-induced oxidative damage in HepG-2 cells by rice protein hydrolysates pretreated with electron beams.

In this paper, electron beam irradiated rice protein hydrolysates (ERPHs) were assessed for their ability to prevent hydrogen peroxide-induced oxidative stress in human HepG-2 cells. The related mechanism was also studied by analyzing the structural changes. Cytotoxicity experiments showed that rice protein hydrolysates pretreated with electron beam irradiation (EBI) were not toxic to cells if appropriate concentrations were applied. Cell viability markedly increased when the cells were treated with ERPHs before H2O2 induction. Furthermore, the ERPHs effectively suppressed H2O2-induced ROS production and lipid peroxidation and increased the protein expression levels of the intracellular antioxidant enzymes SOD, GSH-Px and CAT in H2O2-stressed HepG-2 cells. Consequently, the loss of mitochondrial membrane potential and cell apoptosis was alleviated. Circular dichroism analysis showed that pretreatment of rice protein with EBI significantly changed the secondary structure (the conversion of α-helices to random coils), which is beneficial to the improvement of its antioxidative activity. ERPHs exhibited stronger antioxidative effects than those without irradiation, possibly because of the difference in molecular weight distribution and amino acid composition. These findings indicate an efficient way to produce peptides with better antioxidant activity.

Effect of high energy electron beam on proteolysis and antioxidant activity of rice proteins.

This research focused on the effects of electron beam irradiation (EBI) on the hydrolysis and antioxidant activity of rice proteins (RPs). The RPs were treated with 0, 5, 10, 20 and 30 kGy doses of EBI. The results showed that EBI pretreatment improved significantly (P < 0.05) the degree of hydrolysis, increasing the DH value by more than 15.09% at a dose of 30 kGy. In addition, radical scavenging results showed that EBI treatment had effects on antioxidant activity and could increase the DPPH and ABTS+ radical scavenging activity of rice protein hydrolysates (RPHs) by 32.06% and 79.11%, respectively (30 kGy). The CAA test also confirmed that EBI pretreatment could effectively improve the ability of RPHs to remove intracellular free radicals. Scanning electron microscopy indicated that EBI treatment destroyed microstructures and resulted in cracks and fragments of RPs. Circular dichroism analysis showed that EBI affected the secondary structure of RPs by destroying the α-helix structure. Changes in the UV visible spectra indicated unfolding of RPs by EBI. Amino acid and molecular weight distribution analysis revealed that EBI pretreatment could increase the ratio of antioxidant-related amino acids and produce smaller peptides. Therefore, EBI pretreatment is an efficient method to promote protein proteolysis due to its effect on the molecular conformation as well as on protein microstructure. Moreover, EBI treatment applied before enzymatic hydrolysis has the potential to prepare hydrolysates with high bioactivity.

Decitabine Compared With Conventional Regimens in Older Patients With Acute Myeloid Leukemia: A Meta-Analysis.

BACKGROUND: The incidence of acute myeloid leukemia (AML) increases with age. The overall prognosis remains poor for older patients. Studies on the efficacy of decitabine, an epigenetic agent, in older patients with AML have reported conflicting results. MATERIALS AND METHODS: For this meta-analysis, we performed a literature search and collected 38 studies (including 3298 patients with AML) to evaluate the role of decitabine in elderly patients with AML. We used complete response (CR) or overall response (OR) rate as indicators of effectiveness. RESULTS: Patients treated with decitabine have a higher CR/OR rate than those treated with low-dose cytarabine (CR, 2.60; 95% confidence interval [CI], 1.64-4.14; OR, 4.88; 95% CI, 1.98-12.04) or CAG/HAG (low-dose epirubicin and cytarabine with granulocyte stimulating factor/low-dose homoharringtonine and cytarabine with granulocyte stimulating factor) regimens (CR, 2.53; 95% CI, 1.98-3.23; OR, 2.89; 95% CI, 2.24-3.73). However, patients treated with decitabine had a CR rate equivalent to those treated with intensive chemotherapy (CR, 0.58; 95% CI, 0.28-1.22; P = .15). Use of decitabine in combination with other regimens resulted in a higher CR/OR rate than did use of decitabine alone (P < .001); there was no significant difference in infection rates and early death rates (P > .05). CONCLUSION: The findings presented in this article show that decitabine is effective and safe for the treatment of older patients with AML.

Silence of lncRNA ANRIL represses cell growth and promotes apoptosis in retinoblastoma cells through regulating miR-99a and c-Myc.

Retinoblastoma is a rare cancer of the immature retina. This study designed to see the function of the lncRNA ANRIL in retinoblastoma Y79 cells. ANRIL, miR-99a and c-Myc expression in Y79 cells was altered by transfection and then trypan blue, transwell assay and flow cytometry were carried out to evaluate the changes of cell phenotype. The connection between ANRIL, miR-99a and c-Myc was measured by luciferase reporter assay and RNA immunoprecipitation analysis. As a result, ANRIL expression was highly expressed in human retinoblastoma tissue as relative to the adjacent noncancerous tissues. ANRIL suppression inhibited Y79 cells viability, migration, invasion, while promoted apoptosis. ANRIL negatively regulated miR-99a by binding to miR-99a. Silence of miR-99a reversed the ANRIL-knockdown effects on Y79 cells. miR-99a overexpression suppressed Y79 cell viability, migration, invasion, and enhanced apoptosis through downregulating c-Myc. Meanwhile, we found that miR-99a inhibited JAK/STAT and PI3K/AKT pathways. To conclude, it seems that ANRIL suppression inhibits cell growth and metastasis in retinoblastoma Y79 cells by regulating miR-99a and c-Myc.

Effect of insulin on thyroid cell proliferation, tumor cell migration, and potentially related mechanisms.

BACKGROUND: Diabetes has recently been identified as a risk factor for a variety of cancers, possibly due to hyperinsulinemia or exogenous insulin use. Thyroid cancer is the most common endocrine malignancy, and its incidence has been exponentially increasing worldwide at an alarming rate. The aim of this study was to establish whether insulin use affects thyroid cancer development and progression, specifically cell proliferation and migration in vitro. METHODS: In this study, we investigated the effects of the insulin agents most commonly used in the clinic, regular human insulin (HI) and insulin glargine (IG), on the proliferation and migration of thyroid cells. RESULTS: Both HI and IG affected the thyroid cells in a dose-dependent manner and at high concentrations significantly promoted thyroid cell proliferation and tumor cell migration. The promoting effect might be elicited by activation of the insulin receptor and insulin-like growth factor-1 receptor and through the downstream Akt-signaling pathway, which inhibits the activity of the tumor-suppressor FoxO3a. In particular, MAPK-signaling cascades were activated in papillary thyroid carcinoma cell-1 cells but not in follicular rat thyroid-5 cells. CONCLUSION: The in vitro evidence demonstrated that HI and IG can promote thyroid cell proliferation and tumor cell migration at supraphysiological concentrations, but the effect was not significant at low concentrations. Whether high-dose insulins could affect diabetic patients with thyroid cancer or undetected (pre)cancerous lesions needs further in vivo study. ABBREVIATIONS: HI: human regular insulin; IG: insulin glargine; IR: insulin receptor; IGF-1R: insulin-like growth factor-1 receptor; Akt: protein kinase B (PKB); MAPK: mitogen-activated protein kinase; FoxO3a: the forkhead box-containing protein: class O 3a.

Wilms' tumor 1-associating protein plays an aggressive role in diffuse large B-cell lymphoma and forms a complex with BCL6 via Hsp90.

BACKGROUND: Wilms' tumor 1-associating protein (WTAP) is a nuclear protein, which is ubiquitously expressed in many tissues. Furthermore, in various types of malignancies WTAP is overexpressed and plays a role as an oncogene. The function of WTAP in diffuse large B-cell lymphoma (DLBCL), however, remains unclear. METHODS: Immunohistochemistry was applied to evaluate the levels of WTAP expression in DLBCL tissues and normal lymphoid tissues. Overexpression and knock-down of WTAP in DLBCL cell lines, verified on mRNA and protein level served to analyze cell proliferation and apoptosis in DLBCL cell lines by flow cytometry. Finally, co-immunoprecipitation (Co-IP), IP, and GST-pull down assessed the interaction of WTAP with Heat shock protein 90 (Hsp90) and B-cell lymphoma 6 (BCL6) as well as determined the extend of its ubiquitinylation. RESULTS: WTAP protein levels were consistently upregulated in DLBCL tissues. WTAP promoted DLBCL cell proliferation and improved the ability to confront apoptosis, while knockdown of WTAP in DLBCL cell lines allowed a significant higher apoptosis rate after treatment with Etoposide, an anti-tumor drug. The stable expression of WTAP was depended on Hsp90. In line, we demonstrated that WTAP could form a complex with BCL6 via Hsp90 in vivo and in vitro. CONCLUSION: WTAP is highly expressed in DLBCL, promoting growth and anti-apoptosis in DLBCL cell lines. WTAP is a client protein of Hsp90 and can appear in a complex with BCL6 and Hsp90 in DLBCL. Down-regulation of WTAP could improve the chemotherapeutic treatments in DLBCL.

A polysaccharide from green tea (Camellia sinensis L.) protects human retinal endothelial cells against hydrogen peroxide-induced oxidative injury and apoptosis.

Oxidative damage of retinal pigment epithelium (RPE) cells is involved in the pathogenesis age related macular degeneration (AMD). The purpose of this study was to evaluate the potential protective effect of a purified green tea polysaccharide (GTWP) against hydrogen peroxide (H2O2) induced oxidative stress and apoptosis in human retinal pigment epithelial cells (ARPE-19 cells). Human ARPE-19 cells were treated with 1 h of 500 µM H2O2 before incubation with GTWP for 24 h. Pretreatment of GTWP decreased H2O2-induced cell death and cell apoptosis, and efficiently suppressed the intracellular ROS production and malondialdehyde (MDA) generation induced by H2O2 treatment. Moreover, a loss of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione (GSH) activities were restored to normal level in H2O2-induced ARPE-19 cells upon GTWP (100 µg/ml) exposure. Also, the tendency of increased protein expression of Bax and cleaved-caspsae-3, as well as decrease of Bcl-2 protein in ARPE-19 cells challenged with H2O2 was changed to individual opposite way, thus inhibiting the apoptotic cell death. Our results demonstrated that GTWP protected RPE cells against oxidative injury through activation of anti-apoptotic and endogenous antioxidant enzymes signaling pathway, suggesting GTWP has attractive therapeutic potential to AMD.

LITAF is a potential tumor suppressor in pancreatic cancer.

Early diagnosis of pancreatic cancer, one of the most deadly cancers with low survival rates, is difficult, and effective biomarkers are urgently needed. Lipopolysaccharide-induced tumor necrosis factor-α factor (LITAF) has been recently proposed as a potential tumor suppressor gene in several types of cancer. Here, we analyzed the biological function of LITAF in pancreatic cancer. The LITAF gene and protein levels were decreased in pancreatic tumor tissues compared with their paired adjacent non-cancerous tissues. In addition, patients with the lower LITAF protein expression had lower disease-free survival rates. The decreased LITAF expression correlated with LITAF promoter hypermethylation in pancreatic cancer cells and tissues. Moreover, promoter demethylation dose-dependently increased the LITAF transcription. Importantly, LITAF demethylation suppressed proliferation and cell cycle progression, and enhanced apoptosis of pancreatic cancer cells. Together, our results indicate that LITAF functions as a tumor suppressor gene in pancreatic cancer cells, and might serve as a novel biomarker for early diagnosis of pancreatic cancer.

Isolation of a novel calcium-binding peptide from wheat germ protein hydrolysates and the prediction for its mechanism of combination.

To isolate a novel peptide with specific calcium-binding capacity, wheat germ protein was hydrolyzed. The hydrolysates were purified using ultrafiltration, anion-exchange chromatography, gel filtration chromatography, and reversed-phase high performance liquid chromatography. The amino acid sequence of the purified peptide was determined and confirmed to be FVDVT (Phe-Val-Asp-Val-Thr). The calcium-binding capacity of FVDVT reached 89.94±0.75%, increased by 86.37% compared to the hydrolysates. The chelating mechanism between FVDVT and calcium was further investigated by Ultraviolet-Visible absorption spectroscopy, Fourier transform infrared spectroscopy, X-ray diffraction, and 1H nuclear magnetic resonances spectroscopy. The results indicated that the oxygen atoms of the carboxy group and the nitrogen atoms of the amido group provided major binding sites. In addition, aspartic acid and threonine show considerable capacity for incorporating with calcium by donating electron pairs. This study provides a feasible approach to isolate calcium-binding peptides and to clarify the possible binding mechanism of calcium and peptide.

The feedback loop of LITAF and BCL6 is involved in regulating apoptosis in B cell non-Hodgkin's-lymphoma.

Dysregulation of the apoptotic pathway is widely recognized as a key step in lymphomagenesis. Notably, LITAF was initially identified as a p53-inducible gene, subsequently implicated as a tumor suppressor. Our previous study also showed LITAF to be methylated in 89.5% B-NHL samples. Conversely, deregulated expression of BCL6 is a pathogenic event in many lymphomas. Interestingly, our study found an oppositional expression of LITAF and BCL6 in B-NHL. In addition, LITAF was recently identified as a novel target gene of BCL6. Therefore, we sought to explore the feedback loop between LITAF and BCL6 in B-NHL. Here, our data for the first time show that LITAF can repress expression of BCL6 by binding to Region A (-87 to +65) containing a putative LITAF-binding motif (CTCCC) within the BCL6 promoter. Furthermore, the regulation of BCL6 targets ( PRDM1 or c-Myc) by LITAF may be associated with B-cell differentiation. Results also demonstrate that ectopic expression of LITAF induces cell apoptosis, activated by releasing cytochrome c, cleaving PARP and caspase 3 in B-NHL cells whereas knockdown of LITAF robustly protected cells from apoptosis. Interestingly, BCL6, in turn, could reverse cell apoptosis mediated by LITAF. Collectively, our findings provide a novel apoptotic regulatory pathway in which LITAF, as a transcription factor, inhibits the expression of BCL6, which leads to activation of the intrinsic mitochondrial pathway and tumor apoptosis. Our study is expected to provide a possible biomarker as well as a target for clinical therapies to promote tumor cell apoptosis.

Protective effects of rice dreg protein hydrolysates against hydrogen peroxide-induced oxidative stress in HepG-2 cells.

In this paper, the effects of rice dreg protein hydrolysates (RDPHs) obtained by various proteases on hydrogen peroxide-induced oxidative stress in HepG-2 cells were investigated. Cell cytotoxicity was evaluated through the aspects of cell viability, ROS level, antioxidant enzyme activity, and production of malondialdehyde (MDA). Cell apoptosis was assessed by flow cytometry. Molecular weight distribution was analyzed by gel permeation chromatography, and amino acid composition was measured using an automatic amino acid analyzer. The survival of cells and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were significantly increased through the pre-incubation of HepG-2 cells with RDPHs before H2O2 exposure. Additionally, these pretreatments also resulted in a reduction in ROS and MDA levels. As a result, apoptosis and loss of mitochondrial membrane potential of the HepG-2 cells were alleviated. Furthermore, the protective effects of protein hydrolysates obtained by various proteases were noticeably distinct, in which RDPHs prepared by alkaline protease showed higher antioxidant activities. The difference in the protective effects might be attributed to the specific peptide or amino acid composition. Therefore, enzymatic hydrolysis with different enzymes studied here could attenuate H2O2-induced cell damage, and the type of protease greatly influenced the anti-oxidative activity. Particularly, optimum use of Alcalase could produce peptides with higher antioxidant activity.