A review of endogenous non-starch components in cereal matrix: spatial distribution and mechanisms for inhibiting starch digestion.

As compared with exogenous components, non-starch components (NSCS), such as proteins, lipids, non-starch polysaccharides (NSPs), and polyphenols, inherently present in cereals, are more effective at inhibiting starch digestibility. Existing research has mostly focused on complex systems but overlooked the analysis of the in-situ role of the NSCS. This study reviews the crucial mechanisms by which endogenous NSCS inhibit starch digestion, emphasizing the spatial distribution-function relationship. Starch granules are filled with pores/channels-associated proteins and lipids, embedding in the protein matrix, and maintained by endosperm cell walls. The potential starch digestion inhibition of endogenous NSCS is achieved by altering starch gelatinization, molecular structure, digestive enzyme activity, and accessibility. Starch gelatinization is constrained by endogenous NSCS, particularly cell wall NSPs and matrix proteins. The stability of the starch crystal structure is enhanced by the proteins and lipids distributed in the starch granule pores and channels. Endogenous polyphenols greatly inhibit digestive enzymes and participate in the cross-linking of NSPs in the cell wall space, which together constitute a physical barrier that hinders amylase diffusion. Additionally, the spatial entanglement of NSCS and starch under heat and non-heat processing conditions reduces starch accessibility. This review provides novel evidence for the health benefits of whole cereals.

Effects of foliar selenium application on Se accumulation, elements uptake, nutrition quality, sensory quality and antioxidant response in summer-autumn tea.

Summer-autumn tea is characterized by high polyphenol content and low amino acid content, resulting in bitter and astringent teast. However, these qualities often lead to low economic benefits, ultimately resulting in a wastage of tea resources. The study focused on evaluating the effects of foliar spraying of glucosamine selenium (GLN-Se) on summer-autumn tea. This foliar fertilizer was applied to tea leaves to assess its impact on plant development, nutritional quality, elemental uptake, organoleptic quality, and antioxidant responses. The results revealed that GlcN-Se enhanced photosynthesis and yield by improving the antioxidant system. Additionally, the concentration of GlcN-Se positively correlated with the total and organic selenium contents in tea. The foliar application of GlcN-Se reduced toxic heavy metal content and increased the levels of macronutrients and micronutrients, which facilitated adaptation to environmental changes and abiotic stresses. Furthermore, GlcN-Se significantly improved both non-volatile and volatile components of tea leaves, resulting in a sweet aftertaste and nectar aroma in the tea soup. To conclude, the accurate and rational application of exogenous GlcN-Se can effectively enhance the selenium content and biochemical status of tea. This improvement leads to enhanced nutritional quality and sensory characteristics, making it highly significant for the tea industry.

Study on the Treatment of Tuberous Sclerosis with Rapamycin Nanomicelles Based on Abdominal Ultrasound.

In recent years, mTOR signaling pathway has been found to be the main bridge between TSC1/TSC2 gene mutation and tuberous sclerosis phenotype. Although mTOR inhibitors have been reported to treat tuberous sclerosis in foreign countries, there is still a lack of long-term follow-up results and clinical treatment experience in children. Therefore, research at home and abroad is actively focusing on the mTOR signaling pathway to further clarify the pathogenesis of the disease, and from a clinical point of view, to summarize the clinical data of more patients treated with mTOR inhibitors, to conduct a long-term follow-up and exploration of rapamycin treatment, and to summarize mature treatment experience. This is also the research hotspot of tuberous sclerosis. Based on the study of the treatment of tuberous sclerosis patients with rapamycin nanomicelles by abdominal ultrasound, the therapeutic effect and safety were compared and evaluated through the observation and description of the clinical seizure control and the recovery of EEG peak out of rhythm in children with tuberous sclerosis and infantile spasm.

Expression and protease characterization of a conserved protein YgjD in Vibrio harveyi.

The glycopeptidase GCP and its homologue proteins are conserved and essential for survival of bacteria. The ygjD gene (Glycopeptidase homologue) was cloned from Vibrio harveyi strain SF-1. The gene consisted of 1,017 bp, which encodes a 338 amino acid polypeptide. The nucleotide sequence similarity of the ygjD gene with that of V. harveyi FDAARGOS 107 was 95%. The ygjD gene also showed similarities of 68%, 67% and 50% with those of Salmonella enterica, Escherichia coli and Bacillus cereus. The ygjD gene was expressed in E. coli BL21 (DE3) and the recombinant YgjD was purified by Ni2+ affinity chromatography column. The purified YgjD showed a specific 37 kDa band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and exhibited protease activities of 59,000 units/mg, 53,700 units/mg and 8,100 units/mg, respectively, on N-Acetyl-L-tyrosine ethyl ester monohydrate (ATEE), N-Benzoyl-L-tyrosine ethyl ester (BTEE) and N-Benzoyl-DL-arginine-4-nitroanilide hydrochloride (BAPNA) substrates. When the conserved amino acids of His111, Glu113 and His115 in the YgjD were replaced with alanine, respectively, the protease activities of the mutants were partly decreased. The two conserved His111 and His115 of YgjD were mutated and the protein lost the protease activity, which implied that the two amino acid played very important roles in maintaining its protease activity. The addition of the purified YgjD to the culture medium of V. harveyi strain SF-1 can effectively promote the bacteria growth. These results indicated that the protease activities may be involved in the survival of bacteria.

Establishment and Characterization of a CTC Cell Line from Peripheral Blood of Breast Cancer Patient.

BACKGROUND: Circulating tumor cell (CTC)-based patient-derived cells are ideal models for investigating the molecular basis of cancer. However, the rarity and heterogeneity of CTCs as well as the difficulties of primary culture limit their practical application. Establishing efficient in vitro culture methods and functionally characterizing CTCs is essential for cancer studies. To this end, we developed an experimental protocol for the isolation, expansion, and identification of breast cancer CTCs. METHODS: The CTC-3 cell line was established from peripheral blood cells of a breast cancer patient. A karyotype analysis was performed. The molecular profile was assessed by flow cytometry, quantitative real-time PCR, and western blot. The characteristics of tumors formed by CTC-3 cells were evaluated by cell growth and tumor sphere formation assays and in a mouse xenograft model. The tumors were analyzed by immunohistochemistry, immunofluorescence analysis, and hematoxylin and eosin staining. RESULTS: The CTC-3 cell line showed more aggressive growth both in vitro and in vivo than the widely used MCF-7 breast cancer cell line. CTC-3 cells were also more resistant to chemotherapeutic agents, and gene profiling indicated higher expression levels of the epithelial-to-mesenchymal transition and stemness markers as compared to MCF-7 cells. CONCLUSIONS: CTC-3 cells are a better model for investigating the malignant behavior of breast cancer than existing cell lines.

Synthesis of Zinc oxide nanoparticles from Marsdenia tenacissima inhibits the cell proliferation and induces apoptosis in laryngeal cancer cells (Hep-2).

Biosynthesis of Zinc oxide nanoparticles (ZnONPs) from natural plants stands as a promising nanodrug delivery system in cancer therapeutics. Marsdenia tenacissima (M.t), a Chinese medicinal plant has been extensively used as clinical remedy for treating several types of cancer. In this present study, ZnONPs were synthesized from Marsdenia tenacissima and its anti cancer potency was assessed against in vitro laryngeal cancer cell line Hep-2. The biosynthesized Marsdenia tenacissima Zinc Oxide Nanoparticles [M.t-ZnONPs] was characterized using UV-visible Spec, SEM, TEM and EDAX analysis. The cytotoxic and apoptotic inducing potential of M.t-ZnONPs was assessed by MTT assay and staining such as DCFH-DA, AO/EtBr, Rhodamine 123, DAPI and comet assay. The anticancer potential of M.t-ZnONPs was analysed by Real time PCR analysis of proapoptotic, antiapoptotic and caspases proteins. Our present findings showed characteristic and morphological representation of synthesized M.t-ZnONPs by UV-visible Spec, SEM, TEM and EDAX analysis. M.t-ZnONPs exhibits its cytotoxicity by inhibiting the viability of Hep-2 cells and IC50 value was obtained by MTT assay. The results of apoptotic staining techniques in M.t-ZnONPs treated Hep-2 cells confirm with excess ROS generation, disruption of mitochondrial membrane potential and nuclear damage. The apoptotic inducing potential of M.t-ZnONPs was also evidenced by upregulation of proapoptotic proteins Bax, Caspase 3 & 9 and downregultion of antiapoptotic protein Bcl-2 by RT-PCR analysis. Finally, these results suggested that biosynthesized M.t-ZnONPs is an effective anticancer agent which induces apoptosis in Hep-2 laryngeal cell line and thus conclude that M.t-ZnONPs, a valid anticancer strategy in treating various cancer.

LIN28A-stabilized FBXL19-AS1 promotes breast cancer migration, invasion and EMT by regulating WDR66.

Breast cancer ranks as the top reason for the oncologic mortality for female around the world. The occurrence rate of breast cancer is rapidly rising, especially in China. Although the therapeutic regimes for breast cancer are diverse, the treatment outcome in patients remains dismal. Long non-coding RNAs have been greatly reported as important participators in cancer progression during the past decades. FBXL19 antisense RNA 1 (FBXL19-AS1) has been identified as a novel oncogene in colorectal cancer recently, but its role in breast cancer remains unknown. Present study attempted to explore the functional role and mechanism of FBXL19-AS1 in breast cancer progression. Expression of FBXL19-AS1, lin-28 homolog A (LIN28A), and WD repeat domain 66 (WDR66) were detected by qPCR and Western blotting. Transwell assay was used to detect cell migration and invasion. RIP assay was used to examine interaction between LIN28A and FBXL19-AS1. First, FBXL19-AS1 was highly expressed in breast cancer cell lines. Loss-of-function assays indicated that FBXL19-AS1 promoted cell migration, invasion, and EMT in breast cancer. Mechanistically, FBXL19-AS1 interacted with and was stabilized by LIN28A, an RNA-binding protein which has been reported to be able to stabilize lncRNAs. Moreover, WDR66 expression was promoted by FBXL19-AS1 at mRNA and protein level. Finally, rescue assays suggested that FBXL19-AS1 promoted migration, invasion, and EMT through regulating WDR66 in breast cancer. Current study proved that LIN28A-stabilized FBXL19-AS1 promoted breast cancer metastasis by regulating WDR66, identifying FBXL19-AS1 as a new biological marker in breast cancer.

LncRNA FOXD2-AS1 accelerates the papillary thyroid cancer progression through regulating the miR-485-5p/KLK7 axis.

It has been proved that long noncoding RNAs (lncRNAs) are important modulators in the tumorigenesis and progression of various malignant tumors. Recently, lncRNA FOXD2-AS1 has been reported to be an oncogene in several kinds of human cancers. However, the function of FOXD2-AS1 in papillary thyroid cancer (PTC) has not been well investigated. This study aims to explore the biological role and mechanism of FOXD2-AS1 in PTC. At first, the expression of FOXD2-AS1 was examined in PTC tissues and cell lines with quantitative reverse transcription-polymerase chain reaction (qRT-PCR). FOXD2-AS1 was found to observably upregulated in PTC tissues and cell lines. Kaplan-Meier survival analysis revealed that high expression of FOXD2-AS1 was closely correlated with the unfavorable prognosis of patients with PTC. Based on the TCGA data set, KLK7 was overexpressed in PTC tumor samples. Our experimental data further validated the upregulation of KLK7 in PTC tissues and cell lines. Similarly, high level of KLKF was associated with poor prognosis of patients with PTC. The positive expression association between FOXD2-AS1 and KLK7 was analyzed with Pearson correlation coefficient. Loss-of-function assays revealed that knockdown of FOXD2-AS1 or KLK7 greatly inhibited PTC cell proliferation and migration, while induced cell apoptosis. Results of mechanism experiments suggested that FOXD2-AS1 functioned as a competing endogenous RNA (ceRNA) to enhance the expression of KLK7 by sponging miR-485-5p in PTC. Rescue assays were conducted to verify the function of FOXD2-AS1/miR-485-5p/KLK7 axis in PTC progression.

Decreased expression of TRIM21 indicates unfavorable outcome and promotes cell growth in breast cancer.

BACKGROUND: Tripartite motif-containing protein 21 (TRIM21), an E3 ubiquitin ligase, has been implicated in autoimmune diseases. Dysregulation of TRIM21 contributes to the progression of human malignancies, but its role and clinical significance in breast cancer remain unclear. METHODS: The expression of TRIM21 was examined by quantitative real-time PCR, Western blot, and immunohistochemistry. The role of TRIM21 in the progression of breast cancer was determined using in vitro and in vivo models. The upstream regulation of TRIM21 was investigated by luciferase reporter assay. RESULTS: Here, we showed that TRIM21 expression in breast cancer tissues was decreased at both the mRNA and protein levels in comparison to that in nontumorous tissues. TRIM21 expression was closely associated with tumor size, estrogen receptor, human epidermal growth factor receptor 2, and clinical stage. Low TRIM21 expression was correlated with poor overall and disease-free survival in two independent cohorts containing 1,219 patients with breast cancer. A multivariate Cox regression model suggested TRIM21 as an independent factor for overall survival. In vitro data revealed that TRIM21 expression was suppressed by miR-494-3p directly targeting the 3' untranslated region of TRIM21. Overexpression of TRIM21 impeded cell proliferation and tumor growth in breast cancer, whereas TRIM21 depletion enhanced these capacities. CONCLUSION: Collectively, our findings indicate that TRIM21 serves as a potential prognostic biomarker and functions as a tumor suppressor in breast cancer.

miR-625 suppresses cell proliferation and migration by targeting HMGA1 in breast cancer.

Dysregulation of microRNA contributes to the high incidence and mortality of breast cancer. Here, we show that miR-625 was frequently down-regulated in breast cancer. Decrease of miR-625 was closely associated with estrogen receptor (P = 0.004), human epidermal growth factor receptor 2 (P = 0.003) and clinical stage (P = 0.001). Kaplan-Meier and multivariate analyses indicated miR-625 as an independent factor for unfavorable prognosis (hazard ratio = 2.654, 95% confident interval: 1.300-5.382, P = 0.007). Re-expression of miR-625 impeded, whereas knockdown of miR-625 enhanced cell viabilities and migration abilities in breast cancer cells. HMGA1 was confirmed as a direct target of miR-625. The expressions of HMGA1 mRNA and protein were induced by miR-625 mimics, but reduced by miR-625 inhibitor. Re-introduction of HMGA1 in cells expressing miR-625 distinctly abrogated miR-625-mediated inhibition of cell growth. Taken together, our data demonstrate that miR-625 suppresses cell proliferation and migration by targeting HMGA1 and suggest miR-625 as a promising prognostic biomarker and a potential therapeutic target for breast cancer.