KDM4A-AS1 Promotes Cell Proliferation, Migration, and Invasion via the miR-4306/STX6 Axis in Hepatocellular Carcinoma.

As a primary liver malignancy, hepatocellular carcinoma (HCC) is commonly induced by chronic liver disease and cirrhosis. Bioinformatics analysis reveals that long noncoding RNA KDM4A antisense RNA 1 (KDM4A-AS1) may be aberrantly expressed in HCC and its abnormal expression might influence prognosis in patients. We conducted this study to illustrate the functions and mechanism of KDM4A-AS1 in regulating HCC malignant cell behavior. KD-M4A-AS1, microRNA (miR)-4306 and messenger RNA syntaxin 6 (STX6) expression was examined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). HCC cell proliferation, apoptosis, migration, and invasion were measured by colony forming assays, flow cytometry, wound healing and Transwell assays. The interaction between genes was verified by RNA immunoprecipitation and luciferase reporter assays. Western blotting was performed to quantify protein expression of STX6 or apoptotic markers. KDM4A-AS1 was highly expressed in HCC cells and tissues. KDM4A-AS1 knockdown led to enhanced HCC cell apoptosis and suppressed HCC cell proliferation, migration, and invasion. MiR-4306 bound to and negatively regulated STX6. KDM4A-AS1 directly bound to miR-4306 and thus up-regulated STX6. STX6 overexpression reversed the inhibitory influence of KDM4A-AS1 depletion on HCC malignant behavior. KDM4A-AS1 promotes HCC cell migration, invasion, and growth by upregulating STX6 via miR-4306.

A new high-resolution global topographic factor dataset calculated based on SRTM.

Topography is an important factor affecting soil erosion and is measured as a combination of the slope length and slope steepness (LS-factor) in erosion models, like the Chinese Soil Loss Equation. However, global high-resolution LS-factor datasets have rarely been published. Challenges arise when attempting to extract the LS-factor on a global scale. Furthermore, existing LS-factor estimation methods necessitate projecting data from a spherical trapezoidal grid to a planar rectangle, resulting in grid size errors and high time complexity. Here, we present a global 1-arcsec resolution LS-factor dataset (DS-LS-GS1) with an improved method for estimating the LS-factor without projection conversion (LS-WPC), and we integrate it into a software tool (LS-TOOL). Validation of the Himmelblau-Orlandini mathematical surface shows that errors are less than 1%. We assess the LS-WPC method on 20 regions encompassing 5 landform types, and R2 of LS-factor are 0.82, 0.82, 0.83, 0.83, and 0.84. Moreover, the computational efficiency can be enhanced by up to 25.52%. DS-LS-GS1 can be used as high-quality input data for global soil erosion assessment.

ABIN1 Inhibits Inflammation through Necroptosis-Dependent Pathway in Ulcerative Colitis.

Background: Ulcerative colitis (UC) is characterized by chronic, recurrent intestinal inflammation and intestinal epithelial injury including a wide range of epithelial cell death, ulcers, crypt abscesses, and the formation of fibrosis. The intestinal barrier dysfunction runs through the whole process of the occurrence and development of UC. A recent study revealed that an ubiquitin binding protein ABIN1 played a role in tissue homeostasis and autoimmunity diseases which involved in the anti-inflammatory response of intestinal epithelia cells. However, the roles of ABIN1 in ulcerative colitis pathogenesis remain unclear. Methods: The mRNA and protein expression level of ABIN1 and necroptosis-associated genes (RIPK1, RIPK3, and MLKL) were conducted to investigate the relationship between ABIN1 and necroptosis in clinical UC specimens. Subsequently, the dextran sodium sulfate (DSS)-induced mice colitis model was used to verify the ABIN1 function in vivo. Furthermore, we established ABIN1 gain and loss function assay in CACO-2 to confirm the mechanism in UC in vitro. Results: We found that ABIN1, RIPK1, RIPK3, and MLKL were upregulated in UC sample and DSS-induced colitis. Upon TNF-α stimulation in the intestinal epithelia cell line, overexpression of ABIN1 significantly inhibits necroptosis in the intestinal inflammation model along with the reduction expression of pro-inflammatory cytokines such as IL1B, IL6, IL8, and TNF-α. Blocking RIPK1 by Nec-1s in vivo and in vitro dramatically alleviated the colitis and cell death which shares the same phenotype with ABIN1 overexpression. Conclusion: Hence, the dysregulation of ABIN1 may relate to the uncontrolled necroptosis and inflammation in UC, and negatively regulate the occurrence and process of ulcerative colitis. ABIN1 activation may be considered a therapeutic strategy for UC.

Incorporation of gelatin and Fe2+ increases the pH-sensitivity of zein-anthocyanin complex films used for milk spoilage detection.

In this study, blueberry anthocyanins, gelatin and Fe2+ were incorporated into zein matrix via electrospinning method to prepare colorimetric indicator films for monitoring milk freshness. Gelatin and Fe2+ were incorporated into the film to improve visual discrimination of indicator films' color changes in milk with different freshness degrees and in solution with pH 3-7. Results of SEM, FT-IR and XRD showed that there were intermolecular hydrogen bonds among components, which associated with the larger color difference of indicator films. UV-vis spectral analysis showed that blueberry anthocyanin solutions containing both gelatin and Fe2+ displayed the highest intensity absorption peaks. The optimal ability to distinguish the pH (3-7) of solutions was presented by the indicator film incorporating gelatin (1% (w/v)) and Fe2+ (0.07 mg/mL). Gelatin and Fe2+ increased the color-responsive sensitivity of the indicator film to pH. The film could be successfully used to detect the freshness of milk, whose color changes were visually perceivable: from purple black (fresh milk) to royal purple (spoiling milk) and then to violet red (spoiled milk). The color parameters (L*, a*, R, G and B) of the film revealed a high correlation with the pH/acidity of the milk during storage. The successful application of the indicator film embedding gelatin and Fe2+ for monitoring milk quality changes indicated that the addition of special substances could provide great potential for monitoring freshness and preparing intelligent packaging of food.

Plant nitrogen nutrition: The roles of arbuscular mycorrhizal fungi.

Nitrogen (N) is the most abundant mineral nutrient required by plants, and crop productivity depends heavily on N fertilization in many soils. Production and application of N fertilizers consume huge amounts of energy and substantially increase the costs of agricultural production. Excess N compounds released from agricultural systems are also detrimental to the environment. Thus, increasing plant N uptake efficiency is essential for the development of sustainable agriculture. Arbuscular mycorrhizal (AM) fungi are beneficial symbionts of most terrestrial plants that facilitate plant nutrient uptake and increase host resistance to diverse environmental stresses. AM association is an endosymbiotic process that relies on the differentiation of both host plant roots and AM fungi to create novel contact interfaces within the cells of plant roots. AM plants have two pathways for nutrient uptake: either direct uptake via the root hairs and root epidermis, or indirectly through AM fungal hyphae into root cortical cells. Over the last few years, great progress has been made in deciphering the molecular mechanisms underlying the AM-mediated modulation of nutrient uptake processes, and a growing number of fungal and plant genes responsible for the uptake of nutrients from soil or transfer across the fungi-root interface have been identified. Here, we mainly summarize the recent advances in N uptake, assimilation, and translocation in AM symbiosis, and also discuss how N interplays with C and P in modulating AM development, as well as the synergies between AM fungi and soil microbial communities in N uptake.

Staple food and health: a comparative study of physiology and gut microbiota of mice fed with potato and traditional staple foods (corn, wheat and rice).

The effects of potato and traditional staple foods (corn, wheat and rice) on physiology and gut microbiota were investigated by feeding ICR mice for 12 months. Compared with traditional staple foods, potato significantly improved the food and water intake and survival rate, and inhibited the swelling of viscera of mice, accompanied by a decreased white blood cell count and urine bilirubin content. Furthermore, potato significantly increased the relative abundance of Bacteroides and Faecalibacterium, which are short-chain fatty acid producing bacteria and play very important roles in the maintenance of human health. Meanwhile, potato significantly decreased the relative abundance of spoilage bacteria Pseudomonas and Thiobacillus. Analysis of putative metagenomes indicated that the potato diet upregulated the gene abundance of glycan biosynthesis and metabolism, digestive system and immune system. These findings indicated that potato has the potential to be an excellent substitute for traditional staple foods owing to its good physiological function and favorable gut microbiota modulation.

Metformin alleviates intestinal epithelial barrier damage by inhibiting endoplasmic reticulum stress-induced cell apoptosis in colitis cell model.

To investigate the effect and mechanism of metformin on intestinal epithelial barrier injury in ulcerative colitis. A cell model of colitis was established by co-culture of human colon cancer cell line Caco-2 and human monocyte cell line THP-1. The colitis model cells were treated with metformin at concentration of for Flow cytometry was used to detect Caco-2 cell apoptosis, and Western blotting was used to detect the protein expression of tight junction proteins and endoplasmic reticulum stress-related proteins. After metformin treatment, the apoptosis rate of Caco-2 cells was decreased from (14.22±2.34)% to 0.61)% (=3.119, <0.05), and the expression levels of tight junction protein-1 and claudin-1 increased (=5.172 and 3.546, both <0.05). In addition, the expression levels of endoplasmic reticulum-related proteins glucose regulated protein (GRP) 78, C/EBP homologous protein (CHOP) and caspase-12, as well as the phosphorylation level of PRKR-like endoplasmic reticulum kinase (PERK) and eukaryotic translation initiation factor 2α (eIF2α) decreased (all <0.05). Metformin may alleviate the intestinal epithelial barrier damage in colitis by reducing intestinal epithelial cell apoptosis and increasing the expression of tight junction proteins, which may be associated with the inhibition of endoplasmic reticulum stress-induced apoptotic pathway.

A novel extraction approach and unique physicochemical properties of gelatin from the swim bladder of sturgeon.

BACKGROUND: Gelatin is traditionally produced from mammals and widely applied in the food industry. The production is tedious, time-consuming and environment-unfriendly, while the application is restricted because of zoonosis risk and religious sentiment. RESULTS: Gelatin was extracted by hot water from sturgeon swim bladder after defatting with alcohol and hexane. The yield reached to 94.15% under the optimized conditions of 50 °C, 30 min and 10 mL g-1 . Its amino acid and subunit profiles were similar to type I collagen. Compared to commercial porcine, bovine and piscine gelatins, it exhibited higher whiteness (3.38), emulsion activity (171.76 m2 g-1 ), gel strength (853.23 g), water-holding capacity (92.37%) and viscoelasticity (0.03). But the transmittance (40.56% at 450 nm and 59.07% at 620 nm), emulsion stability (30.09 min), foam expansion (203.00) and stability (26.92), gelling (16.88 °C) and melting temperature (21.80 °C) were lower. While the pH (6.87) and viscosity (28.60 mPa s) were moderate. Moreover, it made better hydrogels and nanofibers. CONCLUSION: Gelatin was extracted from sturgeon swim bladder using a clean and efficient approach, and exhibited unique properties and great potential for the food industry. © 2020 Society of Chemical Industry.

CircB3GNTL1 and miR-598 regulation effects on proliferation, apoptosis, and glutaminolysis in gastric cancer cells.

This study was performed to research circB3GNTL1 control, miR-598 and its mechanism for cell proliferation, apoptosis and glutamine breakdown. For this purpose, CirB3GNTL1 and miR-598 expressions were detected by qRT-PCR in gastric and cell lines; MTT tests were performed to detect proliferation; flow cytometry was established in flow; glutamine decomposition was evaluated with glutamine, glutamic acid and α-keto-glutaric acid (α-KG) expression; Bcl-2, PCNA, ASCT2 and GLS1 expression levels were calculated; Methods of expression were calculated. The results showed that CircB3GNTL1 expression was up-regulated and miR-598 expression was up-regulated in gastric cancer tissues and cell lines; Knockdown circB3GNTL1 prevented proliferation and glutamine decomposition of the gastric cancer cells and induced apoptosis compared with normal para-cancers and gastric cancer cell lines. Results circB3GNTL1 can target and control miR-598 expression, and miR-598 can reverse the proliferation, apoptosis, and decomposition of glutamine from gastric cancer cells by knockdown circB3GNTL1. It was concluded that CirB3GNTL1 prevents decomposition of glutamines and induces apoptosis by controlling miR-598 in gastric cancer cells.

Functional analysis of the OsNPF4.5 nitrate transporter reveals a conserved mycorrhizal pathway of nitrogen acquisition in plants.

Low availability of nitrogen (N) is often a major limiting factor to crop yield in most nutrient-poor soils. Arbuscular mycorrhizal (AM) fungi are beneficial symbionts of most land plants that enhance plant nutrient uptake, particularly of phosphate. A growing number of reports point to the substantially increased N accumulation in many mycorrhizal plants; however, the contribution of AM symbiosis to plant N nutrition and the mechanisms underlying the AM-mediated N acquisition are still in the early stages of being understood. Here, we report that inoculation with AM fungus Rhizophagus irregularis remarkably promoted rice (Oryza sativa) growth and N acquisition, and about 42% of the overall N acquired by rice roots could be delivered via the symbiotic route under N-NO3- supply condition. Mycorrhizal colonization strongly induced expression of the putative nitrate transporter gene OsNPF4.5 in rice roots, and its orthologs ZmNPF4.5 in Zea mays and SbNPF4.5 in Sorghum bicolor OsNPF4.5 is exclusively expressed in the cells containing arbuscules and displayed a low-affinity NO3- transport activity when expressed in Xenopus laevis oocytes. Moreover, knockout of OsNPF4.5 resulted in a 45% decrease in symbiotic N uptake and a significant reduction in arbuscule incidence when NO3- was supplied as an N source. Based on our results, we propose that the NPF4.5 plays a key role in mycorrhizal NO3- acquisition, a symbiotic N uptake route that might be highly conserved in gramineous species.

[Preliminary study on CASK/Id1 pathway in fibroblasts of human keloid].

OBJECTIVE: To verify the existence and significance of calcium/calmodulin dependent serine protein kinase/inhibitors of differentiation 1 (CASK/Id1) pathway in fibroblasts of human keloid. METHODS: Immunofluorescence laser was used to confirm CASK and Id1 protein expression and localization in fibroblasts of the keloid and normal skin. RT-PCR and Western-blot were adopted to analysis the CASK and Id1 expression and differences between keloid and normal skin fibroblasts. The natural combination of CASK and Id1 protein of keloid fibroblasts was tested by immunoprecipitation. RESULTS: CASK and Id1 protein expression were both found in fibroblast cells of keloid and normal skin under normal circumstances. Most of CASK and Id1 were distributed in the cytoplasm and nucleus of fibroblasts. The results of RT-PCR showed that the expression of CASK mRNA in the keloid group was 0.658 +/- 0.024, which was lower than that in the normal control group (1.076 +/- 0.008, t = 11.159, P < 0.05). The expression of Id1 mRNA was 0.497 +/- 0.014, which was higher than that in the normal control group (0.307 +/- 0.017, t = 15.148, P < 0.05). The results of Western-blot showed that the expression level for CASK protein in the keloid group was 0.057 +/- 0.006, which was lower than that in the normal control group (0.168 +/- 0.012, t = 13.524, P < 0.05); the expression of Id1 protein was 0.812 +/- 0.035, which was higher than that in the normal control group (0.368 +/- 0.031, t = 16.356, P < 0.05). The results of immunoprecipitation showed that Id1 could be detected in the CASK precipitate, while CASK also could be detected in the Id1 precipitate. There was a natural binding of CASK and Id1 in keloid fibroblasts. CONCLUSION: CASK/Id1 signal pathway may be existed and involved in the proliferation of keloid fibroblasts, which is related with the occurrence of keloid.

[Expressions of the related genes in fibroblasts of human keloid and the effect of artesunate].

OBJECTIVE: To verify whether abnormal expression of calcium/calmodulin dependent serine protein kinase (CASK) and inhibitors of differentiation 1 (ID1) exist in Fb of keloid, and to observe the effect of artesunate on two genes. METHODS: Fifteen samples of keloid and 12 samples of normal skin tissue (discarded) excised from patients admitted to our hospital were collected. Tissue particle adherent method was used in the primary culture of Fb, and cells from the third to the eighth passage were used for test. Expressions of CASK and ID1 in Fb harvested from both sources were observed with immunofluorescence staining. Fb of keloid were stimulated with artesunate in various concentration for different time, and the median inhibitory concentration (IC50) was determined with the MTT colorimetric assay, which served as the intervention concentration of artesunate. Fb of normal skin were set as normal control group (NC, treated with medium solution). Fb of keloid were divided into scar control group (SC, treated with medium solution) and scar administration group (SA, treated with artesunate in IC50). The cycle and apoptosis of Fb were detected with flow cytometric assay, and the nucleic acid and protein expressions of CASK and ID1 of Fb in each group were determined with RT-PCR and Western blotting. Data were processed with one-way analysis of variance and LSD-t test. RESULTS: Expressions of CASK and ID1 were detected in two kinds of Fb. The concentration of 75 mg/L was selected as the intervention concentration of artesunate. (1) There were statistically significant differences among the three groups in the percentages of cells in G0/G1 phase and G2/M phase (with F values respectively 118.064 and 163.840, P values all below 0.01). The percentage of cells in G0/G1 phase of group SA was (91.4 ± 1.4)%, which was significantly higher than that of group SC and group NC [respectively (80.7 ± 0.3)% and (82.4 ± 0.6)%, with t values respectively 12.740 and 9.872, P values all below 0.05]. The percentage of cells in G2/M phase of group SA was (6.9 ± 0.3)%, which was significantly lower than that of group SC and group NC [respectively (13.7 ± 0.3)% and (12.7 ± 0.8)%, with t values respectively 43.702 and 12.276, P values all below 0.05]. (2) There were statistically significant differences among the three groups in the early and late apoptotic rates (with F values respectively 61.879 and 4710.862, P values all below 0.01). The early and late apoptotic rates of group SA were respectively (7.1 ± 1.0)% and (14.9 ± 0.3)%, which were significantly higher than those of group SC and group NC [with early apoptotic rate respectively (2.6 ± 0.4)% and (2.7 ± 0.3)%, t values respectively 7.974 and 7.767, P values all below 0.05; with late apoptotic rate respectively (2.3 ± 0.3)% and (2.5 ± 0.4)%, t values respectively 72.882 and 69.792, P values all below 0.05]. (3) The mRNA expression of CASK in group SC was 0.658 ± 0.024, and it was lower than that of group NC (1.076 ± 0.008, t = 28.997, P < 0.01) and group SA (0.855 ± 0.008, t = 13.549, P < 0.01). The protein expression of CASK in group SC was 0.067 ± 0.007, and it was lower than that of group NC (0.179 ± 0.015, t = 12.042, P < 0.01) and group SA (0.132 ± 0.010, t = 9.498, P < 0.01). (4) The mRNA expression of ID1 in group SC was 0.416 ± 0.006, which was higher than that of group NC (0.317 ± 0.020, t = 8.299, P < 0.01) and group SA (0.217 ± 0.009, t = 32.417, P < 0.01). The protein expression of ID1 in group SC was 0.789 ± 0.034, and it was higher than that of group NC (0.366 ± 0.029, t = 16.341, P < 0.01) and group SA (0.114 ± 0.006, t = 33.978, P < 0.01). CONCLUSIONS: It is speculated that CASK and ID1 participate in the proliferation of Fb in keloid. The mechanism of artesunate in inhibiting the proliferation of Fb in keloid may be related to the up-regulation of CASK and down-regulation of ID1.