Effects of different drying methods on physicochemical, textural, flavor, and sensory characteristics of yak jerky.

The work aimed to study the effect of four drying methods, namely constant temperature hot air drying (HD), microwave drying (MD), hot air microwave drying (HMD), and gradient hot air drying (GHD), on quality characteristics of dried yak meat. The analyses of physicochemical, textural, flavor, and sensory characteristics were carried out based on these four drying methods. The results revealed that microwave dried yak jerky exhibited better color and received the highest sensory score. Hardness of samples were affected by the drying methods, which showed significant differences. There were 21 free amino acids (FAAs) detected in dried yak samples. The samples treated by microwave drying showed the highest total free amino acid content (73.30 mg/100 g) and the EUC value was significantly higher than other methods, indicating the sample displayed greater flavor. A total of 153 volatile compounds were identified in dried yak meat samples, primarily including aldehydes, ketones, and esters. Moreover, the sensory evaluation indicated that the drying methods could significantly affect on color, flavor, and overall acceptability of different samples. Microwave drying samples scored higher than other drying methods. Overall, considering aspects of quality, time savings, and energy efficiency, microwave drying of yak jerky emerges as a more satisfactory option. This study could provide important theoretical support for the application of drying methods to improve the quality of yak jerky and enhance production efficiency.

Pleckstrin homology and RhoGEF domain containing G4 (PLEKHG4) leads to the activation of RhoGTPases promoting the malignant phenotypes of thyroid cancer.

Thyroid cancer (TC) is one of the most common endocrine system cancers, and its incidence is elevating. There is an urgent need to develop a deeper understanding of TC pathogenesis and explore new therapeutic target for its treatment. This study aimed to investigate the effects of pleckstrin homology and RhoGEF domain containing G4 (PLEKHG4) on the progression of TC. Herein, 29 pairs of TC and adjacent tissues were used to assess the expression of PLEKHG4. A xenograft model of mouse was established by subcutaneously injected with TC cells. Lung metastasis model was established through left ventricular injection. The results revealed that PLEKHG4 was up-regulated in human TC tissues. PLEKHG4 level was correlated with clinicopathological parameters of TC patients. In vitro assays revealed that PLEKHG4 promoted TC cell proliferation, migration, invasion, and epithelial-mesenchymal transformation. Knockdown of PLEKHG4 led to the opposite effects, and the loss of PLEKHG4 enhanced the apoptosis ability and inhibited the stemness properties of TC cells. These findings were further confirmed by the in vivo growth and lung metastasis of TC tumor. Mechanistically, PLEKHG4 promoted the activation of RhoGTPases RhoA, Cdc42, and Rac1. The inhibitors of these RhoGTPases reversed the PLEKHG4-induced malignant phenotypes. Additionally, ubiquitin-conjugating enzyme E2O (UBE2O), a large E2 ubiquitin-conjugating enzyme acted as an ubiquitin enzyme of PLEKHG4, facilitated its ubiquitination and degradation. In conclusion, PLEKHG4, regulated by UBE2O, promoted the thyroid cancer progression via activating the RhoGTPases pathway. UBE2O/PLEKHG4/RhoGTPases axis is expected to be a novel a therapeutic target for TC treatment.

Umami taste evaluation based on a novel mouse taste receptor cell-based biosensor.

Umami, a taste sensation known for its savory and delicious properties, has garnered considerable attention from both consumers and the food industry. However, current understanding and evaluation of umami characteristics remain limited, presenting a long-standing issue. To address this challenge, we have developed a self-assembled biosensor based on matured taste receptor cells (TRCs), obtained through isolation and culture of taste stem cells. TRCs, as the recognition element, were mounted onto the surface of a glassy carbon electrode (GCE) treated with gold nanoparticles (AuNPs) and poly-L-lysine (PLL). Key parameters including the cell incubation time and concentration were optimized to ensure the optimal performance of the TRCs-based biosensor. AuNPs were deposited onto the GCE surface via 90 s electrochemical reduction. TRCs concentration of 106 cells/mL and incubation time of 12 h were chosen by electrochemical characterization. Using this novel, rapid, and sensitive TRCs-based biosensor, we successfully detected L-monosodium glutamate (MSG) and other umami substances, demonstrating a good linear relationship within the range of 10-9 - 10-5 M between response signals and concentration of MSG stimuli. Our results provide insights into taste signal transduction mechanisms and suggest the potential for biomimetic sensors in intelligent perception applications.

Effects of acetic, malic, and citric acids on the large deformation behaviors of fish gelatin gels.

This research was aimed to quantify the effects of acetic acid, malic acid, and citric acid (0, 0.5, 1.0, and 2.0 g/100 g H2 O) on the stress-strain responses of fish gelatin (FG) gels (2, 4, and 6.67 g/100 g H2 O) under uniaxial compression up to 68% of deformation. The first-order Ogden model fitted quite well for the compression responses of FG gels (R2 = 0.9909-0.9997). Protons from the acids played a key role on weakening the FG gel structures (gel rigidity, µ, decreased 11%-27%), as the µ values and pH values of FG gels were linearly correlated (R2 = 0.8240-0.9748), regardless of the acid type. The addition of an acid also resulted in a significant increase (p < .002) in the strain hardening capacity (α) of gels with 2 g FG/100 g H2 O. Both µ and α values of FG gels with higher gelatin concentrations were less affected by an acid partly due to their stronger buffering effects. The µ and α values of FG gels as affected by acids could not be fully explained based upon the pH changes, implying that the effects of acetate, malate, and citrate ions on the gel structure could not be ignored.

Identification and validation of a CCL18-related signature for prediction of overall survival in patients with uveal melanoma.

Uveal melanoma (UM), the most frequent primary intraocular tumor in adults, has poor prognosis. High C-C motif chemokine ligand 18 (CCL18) has been detected in various tumors and is closely correlated with patients' clinicopathological characteristics. However, the essential role of CCL18 in UM remains unclear. Therefore, this study aimed to explore the prognostic value of CCL18 in UM. Uveal melanoma cells (M17) were transfected with pcDNA3.1-CCL18 si-RNA using Lipofectamine™ 2000. Cell growth and invasion abilities were measured through Cell Counting Kit-8 assay and invasion assay. RNA expression data and clinical and histopathological details were downloaded from the UM in The Cancer Genome Atlas (TCGA-UM) and GSE22138 datasets, which were defined as the training and validation cohorts, respectively. Univariate and multivariate Cox regression analyses were performed to identify significant prognostic biomarkers. The coefficients of these significant biomarkers generated by multivariate Cox proportional hazard regression analysis were used to establish a risk score formula. Functional enrichment analyses were also carried out. We found that downregulated CCL18 inhibits M17 cell growth and invasion in vitro. CCL18 may affect UM progression by altering C-C motif receptor 8 related pathways. Higher CCL18 expression was associated with worse clinical outcomes and tumor-specific death in the TCGA-UM dataset. Based on the coefficients obtained from the Cox proportional hazard regression analysis, a CCL18-related prognostic signature formula was constructed as follows: risk score = 0.05590 × age +2.43437 × chromosome 3 status +0.39496 × ExpressionCCL18. Notably, in this formula, the normal chromosome 3 was coded as 0, whereas the chromosome 3 loss was coded as 1. Each patient was assigned to either low-risk or high-risk groups using the median cut-off in the training cohort. High-risk patients survived for a shorter time than low-risk patients. The time-dependent and multivariate receiver operating characteristic curves showed promising diagnostic efficacy. Multivariate Cox regression analysis demonstrated the potential of this CCL18-related signature as an independent prognostic indicator. These results were validated using the GSE22138 dataset. In addition, in both TCGA-UM and GSE22138 datasets, stratification of clinical correlations and survival analyses based on this signature indicated the involvement of clinical progression and survival outcome in UM. In the high-risk group, Gene Ontology analyses mainly indicated the enrichment of immune response pathways, such as the T cell activation, response to interferon-gamma, antigen processing and presentation, interferon-gamma-mediated signaling pathway, MHC protein complex, MHC class II protein complex, antigen binding, and cytokine binding. Meanwhile, Kyoto Encyclopedia of Genes and Genomes analyses showed enrichments of pathways in cancer, cell adhesion, cytokine-cytokine receptor interaction, chemokine signaling pathway, Th1 and Th2 cell differentiation, and chemokine signaling pathway. Moreover, single-sample gene set enrichment analysis demonstrated the enrichment of almost all immune cells and immune functions in the high-risk group. In summary, a new prognostic CCL18-related signature was successfully established using the TCGA-UM dataset and validated using the GSE22138 dataset with meaningful predictive and diagnostic efficacies. This signature could serve as an independent and promising prognostic biomarker for patients with UM.

A highly sensitive surface-enhanced Raman scattering sensor with MIL-100(Fe)/Au composites for detection of malachite green in fish pond water.

Concerns about food safety have been arisen due to the improper use of chemicals in aquaculture. Malachite green (MG) has attracted attention because of its illegal usage and its potential negative impacts on the environment and public health. Surface-enhanced Raman scattering (SERS) platforms coupled with different SERS substrates have been employed for rapid analysis of MG residues in food. However, the most commonly used SERS substrates were non-reusable and showed limited detection sensitivity. In this study, a novel SERS substrate with a good recyclability and a high sensitivity was prepared by electrostatically assembling together a metal-organic framework material called materials of institute lavoisie-100(Fe) (MIL-100(Fe)) and Au NPs. The lowest detectable concentration of MG was 10-13 M based on the optimal substrate. The SERS sensor was applied for the detection of the trace MG in fish pond water, which was accomplished with the correlation coefficients R2 = 0.991-0.996 in a concentration range of 10-6-10-13 M. Moreover, MIL-100(Fe)/Au was recycled at least five times, realizing a "detection to degradation", showing great potential for food contamination monitoring due to its distinguished performance.

Identification of a prognostic six-immune-gene signature and a nomogram model for uveal melanoma.

BACKGROUND: To identify an immune-related prognostic signature and find potential therapeutic targets for uveal melanoma. METHODS: The RNA-sequencing data obtained from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets. The prognostic six-immune-gene signature was constructed through least absolute shrinkage and selection operator and multi-variate Cox regression analyses. Functional enrichment analysis and single sample GSEA were carried out. In addition, a nomogram model established by integrating clinical variables and this signature risk score was also constructed and evaluated. RESULTS: We obtained 130 prognostic immune genes, and six of them were selected to construct a prognostic signature in the TCGA uveal melanoma dataset. Patients were classified into high-risk and low-risk groups according to a median risk score of this signature. High-risk group patients had poorer overall survival in comparison to the patients in the low-risk group (p < 0.001). These findings were further validated in two external GEO datasets. A nomogram model proved to be a good classifier for uveal melanoma by combining this signature. Both functional enrichment analysis and single sample GSEA analysis verified that this signature was truly correlated with immune system. In addition, in vitro cell experiments results demonstrated the consistent trend of our computational findings. CONCLUSION: Our newly identified six-immune-gene signature and a nomogram model could be used as meaningful prognostic biomarkers, which might provide uveal melanoma patients with individualized clinical prognosis prediction and potential novel treatment targets.

Hypermethylation of microRNA-497-3p contributes to progression of thyroid cancer through activation of PAK1/ß-catenin.

MicroRNA-497 (miR-497) has been reported to be a tumor-suppressive miRNA in thyroid cancer (TC), yet the mechanism is not clearly defined. In this study, we aim to determine the mechanism by which miR-497-3p affects the progression of TC. After characterization of low miR-497-3p expression pattern in TC and normal tissues, we assessed the correlation between miR-497-3p expression and clinicopathological features of TC patients. Its low expression shared associations with advanced tumor stage and lymph node metastasis. ChIP and methylation-specific PCR provided data showing that downregulation of miR-497-3p in TC tissues was induced by DNA methyltransferase-mediated hypermethylation. By performing dual-luciferase reporter assay, we identified that miR-497-3p targeted PAK1 while PAK1 could inhibit ß-catenin expression. Through this mechanism, miR-497-3p exerted the anti-proliferative, anti-invasive, pro-apoptotic, and anti-tumorigenic effects on TC cells on the strength of the results from gain-of-function and rescue experiments. This study suggested that hypermethylation of miR-497-3p resulted in upregulation of ß-catenin dependent on PAK1 and contributed to cancer progression in TC, which highlighted one of miR-mediated tumorigenic mechanism.

Typical Umami Ligand-Induced Binding Interaction and Conformational Change of T1R1-VFT.

Umami taste receptor type 1 member 1/3 (T1R1/T1R3) heterodimer has multiple ligand-binding sites, most of which are located in T1R1-Venus flytrap domain (T1R1-VFT). However, the critical binding process of T1R1-VFT/umami ligands remains largely unknown. Herein, T1R1-VFT was prepared with a sufficient amount and functional activity, and its binding characteristics with typical umami molecules (monosodium l-glutamate, disodium succinate, beefy meaty peptide, and inosine-5'-monophosphate) were explored via multispectroscopic techniques and molecular dynamics simulation. The results showed that, driven mainly by hydrogen bond, van der Waals forces, and electrostatic interactions, T1R1-VFT bound to umami compound at 1:1 (stoichiometric interaction) and formed T1R1-VFT/ligand complex (static fluorescence quenching) with a weak binding affinity (Ka values: 252 ± 19 to 1169 ± 112 M-1). The binding process was spontaneous and exothermic (ΔG, -17.72 to -14.26 kJ mol-1; ΔH, -23.86 to -12.11 kJ mol-1) and induced conformational changes of T1R1-VFT, which was mainly reflected in slight unfolding of α-helix (Δα-helix < 0) and polypeptide chain backbone structure. Meanwhile, the binding of the four ligands stabilized the active conformation of the T1R1-VFT pocket. This work provides insight into the binding interaction between T1R1-VFT/umami ligands and improves understanding of how umami receptor recognizes specific ligand molecules.

A novel umami electrochemical biosensor based on AuNPs@ZIF-8/Ti3C2 MXene immobilized T1R1-VFT.

The bioelectronic tongues based on taste receptors have been emerging with human-like taste perception. However, the practical applications of the receptor-based biosensors were restricted by their narrow and low dynamic ranges. Here, a novel immobilization strategy based on AuNPs@ZIF-8/Ti3C2 MXene was developed to immobilize the umami ligand binding domain (T1R1-VFT), to fabricate an umami biosensor for umami substances detection. Through the synergic effect of AuNPs@ZIF-8 and Ti3C2 MXene, the capacity to load T1R1-VFT was effectively increased, and the response signal was also amplified by approximately 3 times. The proposed biosensor showed an ultrawide dynamic range of 10-11-10-3 M, and a high upper limit of detection, which was closer to the human taste threshold and suitable for detecting foods rich in umami substances. Additionally, the biosensor was successfully applied to detect real samples and analyze the synergistic effects of binary umami substances.

Silencing of histone deacetylase 3 suppresses the development of esophageal squamous cell carcinoma through regulation of miR-494-mediated TGIF1.

BACKGROUND: Deacetylation of histones by histone deacetylase 3 (HDAC3) acts importantly in modulating apoptosis, DNA damage and cellular progression. Herein, we aimed to unravel the functional role of HDAC3 in a lethal disease, esophageal squamous cell carcinoma (ESCC). METHODS: The expression of HDAC3 in clinically collected ESCC tissues was determined by RT-qPCR and immunohistochemistry. As revealed from bioinformatics analysis, the putative relations between HDAC3 and microRNA-494 (miR-494) and between miR-494 and transforming growth factor beta (TGFß)-inducing factor 1 (TGIF1) were further verified by chromatin immunoprecipitation and dual-luciferase reporter gene assay. Functional roles of shRNA-mediated depletion of HDAC3, miR-494 mimic and overexpressed TGIF1 were explored by gain- and loss-of-function assays with regard to ESCC cell biological behaviors. A nude mouse model of ESCC was developed for in vivo validation. RESULTS: HDAC3 was highly expressed in ESCC tissues, suggestive of poor prognosis while TGIF1 was upregulated and miR-494 was downregulated. Mechanistic investigation revealed that HDAC3 inhibited miR-494 expression and TGIF1 was a direct target of miR-494. Furthermore, silencing HDAC3 or overexpressing miR-494 was demonstrated to suppress aggressive phenotypes of ESCC cells both in vitro through the activated TGFß signaling pathway and in vivo, while TGIF1 overexpression induced opposite results. CONCLUSION: Collectively, our findings provided demonstration regarding the oncogenic property of HDAC3 in ESCC via the miR-494/TGIF1/TGFß axis.

Long non-coding ROR promotes the progression of papillary thyroid carcinoma through regulation of the TESC/ALDH1A1/TUBB3/PTEN axis.

Papillary thyroidal carcinoma (PTC) is a common endocrine cancer that plagues people across the world. The potential roles of long non-coding RNAs (lncRNAs) in PTC have gained increasing attention. In this study, we aimed to explore whether lncRNA ROR affects the progression of PTC, with the involvement of tescalcin (TESC)/aldehyde dehydrogenase isoform 1A1 (ALDH1A1)/ßIII-tubulin (TUBB3)/tensin homolog (PTEN) axis. PTC tumor and adjacent tissues were obtained, followed by measurement of lncRNA ROR and TESC, ALDH1A1, and TUBB3 expression. Interactions among lncRNA ROR, TESC, ALDH1A1, TUBB3, and PTEN were evaluated by ChIP assay, RT-qPCR, or western blot analysis. After ectopic expression and depletion experiments in PTC cells, MTT and colony formation assay, Transwell assay, and flow cytometry were performed to detect cell viability and colony formation, cell migration and invasion, and apoptosis, respectively. In addition, xenograft in nude mice was performed to test the effects of lncRNA ROR and PTEN on tumor growth in PTC in vivo. LncRNA ROR, TESC, ALDH1A1, and TUBB3 were highly expressed in PTC tissues and cells. Overexpression of lncRNA ROR activated TESC by inhibiting the G9a recruitment on the promoter of TESC and histone H3-lysine 9me methylation. Moreover, TESC upregulated ALDH1A1 expression to increase TUBB3 expression, which then reduced PTEN expression. Overexpression of lncRNA ROR, TESC, ALDH1A1 or TUBB3 and silencing of PTEN promoted PTC cell viability, colony formation, migration, and invasion while suppressing apoptosis. Moreover, overexpression of lncRNA ROR increased tumor growth by inhibiting PTEN in vivo. Taken together, the current study demonstrated that lncRNA ROR mediated TESC/ALDH1A1/TUBB3/PTEN axis, thereby facilitating the development of PTC.

Taste and stability characteristics of two key umami peptides from pufferfish (Takifugu obscurus).

Takifugu obscurus (T. obscurus) is known for its umami taste. Two taste-active peptides, Pro-Val-Ala-Arg-Met-Cys-Arg (PR-7) and Tyr-Gly-Gly-Thr-Pro-Pro-Phe-Val (YV-8), were proved as key compounds that contributed to the typical taste of T. obscurus. However, whether these peptides have the potential as umami supplements is unknown. The purpose of this study was to investigate the taste characteristics of PR-7 and YV-8, as well as stability at different pH values by sensory evaluation, instrumental analysis and quantum chemical calculation. The results indicated that PR-7 and YV-8 presented umami taste at near neutral pH (6.5-8.0) and had umami-enhancing effects. PR-7 also exhibited significant kokumi activity. Additionally, two peptides showed remarkable stability after different pH treatments, especially YV-8; this may be related to its stable structural property. All the results suggest that both peptides have great potential to be applied in complex foods to provide desirable taste, and act as a feasible alternative to monosodium l-glutamate.

Study on the distribution of umami receptors on the tongue and its signal coding logic based on taste bud biosensor.

Taste signals are uniformly encoded and transmitted to the brain's taste center by taste buds, and the process has not been systematically studied for several decades. The aim of this work was to investigate the distribution of umami receptors on the tongue and its signal coding logic based on the taste bud biosensors. Taste bud biosensors were constructed by immobilizing the taste bud tissues from different tongue regions of the rabbit to the glassy carbon electrode surface; The Shennong information equations were used to analysis the pattern of umami receptors to encode ligands information; The signal amplification capabilities of two types umami receptors (T1R1/T1R3 and mGluRs) were analyzed for the two ligands (L-monosodium glutamate (MSG) and disodium 5'-inosinate (IMP)). The results showed that each taste bud biosensor could sense MSG and IMP with different response currents based on enzyme-substrate kinetics. There was only a small fraction of a great quantity of metabotropic glutamate receptors (mGluRs) could be activated to encode MSG signal. Importantly, T1R1 was more expressed in the rostral tongue cells whose sensitivity to MSG was nearly 100 times stronger than that of caudal tongue cells. The method we proposed made it possible to reveal the distribution and signals coding logic of umami receptors for ligands, which showed great potential to explain the interaction mechanism of umami substances with their receptors more accurately and to develop of artificial intelligent taste sensory.

CircTUBGCP3 facilitates the tumorigenesis of lung adenocarcinoma by sponging miR-885-3p.

BACKGROUND: Circular RNAs (circRNAs) act pivotal roles in the progression of multiple malignancies. However, the underlying mechanisms by which hsa_circ_0007031 (circTUBGCP3) contributes to lung adenocarcinoma (LAC) remain largely unknown. METHODS: The association of circTUBGCP3 expression with clinicopathological characteristics and prognosis in patients with LAC was determined by RT-qPCR and fluorescence in situ hybridization. The in vitro functional experiments as well as a subcutaneous tumorigenesis model were executed to estimate the role of circTUBGCP3 in LAC cells. The interaction between circTUBGCP3 and miR-885-3p was confirmed by RNA immunoprecipitation, luciferase gene report and RT-qPCR assays. The effects of circTUBGCP3 on miR-885-3p-mediated Wnt10b/ß-catenin signaling were evaluated by Western blot. RESULTS: The upregulation of circTUBGCP3 or downregulation of miR-885-3p was associated with the pathological stage and poor survival in patients with LAC. Restored expression of circTUBGCP3 facilitated the growth and invasion of LAC cells, but knockdown of circTUBGCP3 harbored the opposite effects. In mechanism, circTUBGCP3 could act as a sponge of miR-885-3p, which suppressed the cell proliferation and colony formation and attenuated the tumor-promoting effects of circTUBGCP3. Wnt10b as a target of miR-885-3p could be upregulated be circTUBGCP3 and indicate poor survival in patient with LAC. CONCLUSIONS: Our findings demonstrated that circTUBGCP3 promoted LAC progression by sponging miR-885-3p, and might represent a prognostic factor for LAC.

Human-like performance umami electrochemical biosensor by utilizing co-electrodeposition of ligand binding domain T1R1-VFT and Prussian blue.

Over the past decades, due to the desire for artificial umami flavors, apparatuses for detecting the umami taste have constantly been developed. Nevertheless, most information on umami is still acquired through human sensory assessment, which makes it difficult to establish an umami standard or quantify the umami flavor. In this study, the ligand binding domain called venus flytrap (VFT) domain of the umami taste receptor protein T1R1 was used as a recognition element, and an electrochemical biosensor based on a double-signal amplification strategy was constructed using single-walled carbon nanotubes (SWCNTs) and Prussian blue (PB). Moreover, the umami taste of four representative umami substances, inosine-5'-monophosphate (IMP), monosodium L-glutamate (MSG), beefy meaty peptide (BMP), and sodium succinate (WSA), were successfully quantitatively measured using differential pulse voltammetry (DPV) at an electrochemical workstation. Based on an equation (S/N = 3), the low detection limits (LODs) of IMP, MSG, BMP, and WSA were 0.1, 0.1, 0.1, and 0.01 pM, respectively. Meanwhile, a normalized signal intensity of more than 90% was kept for 4 days. The results showed that the biosensor could be used to detect umami substances with high sensitivity and selectivity, and was shown to have human-like performance. To develop the T1R1-VFT biosensor using the above-mentioned method, we utilized the ligand binding domain of the human umami receptor, rather than the entire umami receptor protein, which had a complex structure, having the following advantages: volume reduction, simplicity, and stability. This method has great potential for the detection of umami tastes, instead of using sensory evaluation, and for the development of new artificial flavorings.

Comparison of physicochemical and umami characterization of aqueous and ethanolic Takifugu obscurus muscle extracts.

Most umami substances were developed in aqueous extracts. In this study, we compared the molecular weight distributions and sensory characteristics of ethanol and aqueous Takifugu obscurus muscle extracts, and assessed their taste-related metabolites and peptide profile (<3 kDa) using nuclear magnetic resonance and nano liquid chromatography-mass spectrometry. The potential antioxidant peptide in ethanolic fraction was screened using Peptide Ranker, BIOPEP and quantum chemical simulations. The results indicated that 60% ethanolic extract fraction (60%-F) had the highest umami intensity and more palatable overall taste among all pufferfish extracts. It can be caused by more umami enhancing components such as Asp, Asn, Ala and 5'-AMP, and considerable umami-potential smaller peptides in 60%-F. 60%-F also showed an antioxidant activity, and several antioxidant peptides was screened. The present study indicated the relationship between extract solution and taste characterization, which provided more possibility for the exploitation of umami substances and screening potential activity peptides.

New technology to improve the thermal stability of botulinum toxin type D by biomimetic mineralization.

The advanced biomimetic mineralization technology was applied to protect the Botulinum neurotoxin type D, and the processing of the mineralization granule of botulinum toxin type D was successfully screened. The loss of activity of the toxin protein at different temperatures and the destructive strength of the gastrointestinal tract against the toxin were determined biologically. The lethal toxicity of the mineralized toxin to wild rodents was determined by median lethal dose. Protective tests at different temperatures showed that the preservation period of botulinum toxin type D mineralized sample 2 was significantly higher than that of the control group at three different temperatures, and its toxicity loss was significantly reduced. The damage intensity of the mineralized toxin to the gastrointestinal contents of plateau zokor and plateau pika was significantly reduced. The minimum lethal doses of the mineralized toxin particles to plateau zokor, plateau pika, and mice were 5200, 8,600,000, and 25,000 MLD/kg. These results showed that biomimetic mineralization could greatly improve the thermal stability of botulinum toxin type D and reduce the damaging effect of the gastrointestinal contents of target animals to botulinum toxin type D. The mineralized toxin could be used to control the population density of urban rodents. This research provides new insights into the protection of toxin protein substances.

SOX2-Upregulated microRNA-30e Promotes the Progression of Esophageal Cancer via Regulation of the USP4/SMAD4/CK2 Axis.

Esophageal cancer (EC) is a highly aggressive disease, and its progression involves a complex gene regulation network. Transcription factor SOX2 is amplified in various cancers including EC. A pathway involving SOX2 regulation of microRNAs (miRNAs) and their target genes has been previously revealed. This study aims to delineate the ability of SOX2 to influence the EC progression, with the involvement of miR-30e/USP4/SMAD4/CK2 axis. SOX2 expression was first examined in the clinical tissue samples from 30 EC patients. Effects of SOX2 on proliferation, migration, and invasion alongside tumorigenicity of transfected cells were examined by means of gain- and loss-of-function experiments. EC tissues and cells exhibited high expression of SOX2, miR-30e, and CK2 and poor expression of USP4 and SMAD4. Mechanistically, SOX2 was positively correlated with miR-30e and upregulated the expression of miR-30e. miR-30e specifically targeted USP4, which induced deubiquitination of SMAD4 and promoted its expression. Meanwhile, SMAD4 was enriched in the CK2 promoter region and thus inhibited its expression. SOX2 stimulated EC cell proliferative, invasive, and migratory capacities in vitro and tumor growth in vivo by regulating the miR-30e/USP4/SMAD4/CK2 axis. Collectively, our work reveals a novel SOX2-mediated regulatory network in EC that may be a viable target for EC treatment.

Construction and analysis of the ceRNA network hsa_circ_0031968/miR-3611/GCG in lung adenocarcinoma.

BACKGROUND: A competitive endogenous RNA (ceRNA) network was constructed to examine the potential mechanisms of circular RNAs (circRNAs) in lung adenocarcinoma (LUAD). METHODS: LUAD-related data sets were obtained from the Gene Expression Omnibus (GEO) database and screened for differentially expressed circRNAs (DECs) and differentially expressed microRNAs (DEMs). We identified the target microRNAs (miRNAs) regulated by the DECs and the potential target genes of the miRNA. The basic structure of the DECs were analyzed and enrichment analyses were conducted to determine the function of the circRNA. The Kaplan-Meier method for survival analysis was used to examine the clinical data from The Cancer Genome Atlas (TCGA) database. A protein-protein interaction (PPI) network was constructed to determine the hub genes. The relative expression of the RNA molecules on the ceRNA axis was verified by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analysis. RESULTS: A total of 17 DECs and 237 DEMs were selected for analysis. After reviewing the cancer-specific circRNA database (CSCD), 10 circRNAs were identified. The 432 target miRNAs were screened by circRNA interactome (CRI) and cross-referenced with the DEMs to obtain 126 miRNAs of interest. The expression of miR-3611, which is regulated by hsa_circ_0031968, was found to significantly affect the survival and prognosis of patients with LUAD (P≤0.05). The target gene function of hsa_circ_0031968 was determined to be mainly enriched in SMAD binding, and the signaling pathway was primarily enriched in miRNAs related to cancer. The TCGA database screened out 2,484 differentially expressed mRNAs (DEmRNAs) and intersection analysis with the target gene of miR-3611 revealed 1 gene, namely the proglucagon gene (GCG). Therefore, we chose the hsa_circ_0031968/miR-3611/GCG axis for further research. The expression of GCG was determined to be associated with a poorer survival rate and higher T stage in LUAD patients. Finally, 17 hub genes that interact with GCG were identified. CONCLUSIONS: The ceRNA regulatory network hsa_circ_0031968/miR-3611/GCG was successfully constructed and this provided novel insights into the identification of biomarkers and the pathogenesis of LUAD. This knowledge will contribute to the early diagnosis and development of potential treatment for patients with LUAD.