BrTTG1 regulates seed coat proanthocyanidin formation through a direct interaction with structural gene promoters of flavonoid pathway and glutathione S-transferases in Brassica rapa L.

Introduction: Seed coat color is a significant agronomic trait in horticultural crops such as Brassica rapa which is characterized by brown or yellow seed coat coloration. Previous Brassica rapa studies have shown that BrTTG1 is responsible for seed coat proanthocyanidin formation, which is dependent on the MYB-bHLH-WD40 complex, whereas some studies have reported that TRANSPARENT TESTA GLABRA 1 (TTG1) directly interacts with the structural gene promoters of the flavonoid pathway. Methods: Herein, the brown-seeded inbred B147 and ttg1 yellow-seeded inbred B80 mutants were used as plant materials for gene expression level analysis, gene promoter clone and transient overexpression. Results: The analysis identified eleven structural genes involved in the flavonoid biosynthesis pathway, which are potentially responsible for BrTTG1- dependent seed coat proanthocyanidin formation. The promoters of these genes were cloned and cis-acting elements were identified. Yeast one-hybrid and dual-luciferase assays confirmed that BrTTG1 directly and independently interacted with proCHS-Bra008792, proDFR-Bra027457, proTT12-Bra003361, proTT19-Bra008570, proTT19-Bra023602 and proAHA10-Bra016610. A TTG1-binding motif (RTWWGTRGM) was also identified. Overexpression of TTG1 in the yellow-seed B. rapa inbred induced proanthocyanidin accumulation by increasing the expression levels of related genes. Discussion: Our study unveiled, for the first time, the direct interaction between TTG1 and the promoters of the flavonoid biosynthesis pathway structural genes and glutathione S-transferases in Brassica rapa. Additionally, we have identified a novel TTG1-binding motif, providing a basis for further exploration into the function of TTG1 and the accumulation of proanthocyanidins in seed coats.

[Radix Tetrastigme Polysaccharide Promotes Antitumor Immune Response 
in Lewis Lung Cancer Mice].

BACKGROUND: Lung cancer has a high incidence and mortality rate, but the treatment of lung cancer still lacks low toxicity and efficient anti-tumor drugs. Polysaccharide from radix tetrastigme has development value in anti-tumor treatment methods. This study was to observe the effect of polysaccharide from radix tetrastigme on immune response of Lewis lung cancer mice and explore its molecular mechanism. METHODS: Lewis lung cancer mouse models were established and randomly grouped. The spleen polypeptide group was intragastric with 50 mg/kg spleen polypeptide, and the radix tetrastigme polysaccharide low, medium and high dose groups were intragastric with 62.5, 125 and 250 mg/kg radix tetrastigme polysaccharide, respectively, and the model group and the control group were intragastric with equivolume normal saline. Tumor formation and metastasis were compared. Haematoxylin-eosin (HE) staining was used to observe the pathological changes of tumor cells. Macrophage phagocytosis, apoptosis, M1/M2 polarization, T cell subsets and cytokine levels in peripheral blood were detected by flow cytometry. The proliferation activity of macrophages was detected by methyl thiazolyldiphenyl tetrazolium (MTT) assay. Dendritic cell (DC) antigen presenting function was detected by chlorophenol red-ß-D-galactopyranoside (CPRG) method. Tumor tissue differentiation antigen cluster 47 (CD47) mRNA and protein expression and macrophage signal regulatory protein α (SIRRP α) expression were detected by real time quantitative polymerase chain reaction (RT-qPCR) and Western blot (WB). RESULTS: The tumor inhibition rates and anti-metastasis rates in the 3-dose radix tetrastigme polysaccharide group and the spleen polypeptide group were higher than those in the model group, and the pathological injury of tumor tissue were severer, and the positive rate of phagocytosis of ink by macrophages and the efficiency of phagocytosis of tumor cells were increased; the apoptosis rate of macrophages was decreased; the proliferation activity of macrophages, polarization ratio of macrophages to M1 type, DC antigen presenting ability, CD4+, CD4+/CD8+ levels were increased; the level of serum tumor necrosis factor α (TNF-α), interleukin-1ß (IL-1ß), and the expression of tumor tissue CD47, macrophage SH2-containing protein tyrosine phosphatase 1 (SHP-1), SH2-containing protein tyrosine phosphatase 2 (SHP-2), and phosphorylation signal regulatory protein α (p-SIRPα) were decreased, and the differences were statistically significant (P<0.05). There were no significant differences in the above indexes between low-dose radix tetrastigme polysaccharide group and spleen polypeptide group (P>0.05), and the effects of radix tetrastigme polysaccharide were dose-dependent. CONCLUSIONS: Radix tetrastigme polysaccharide can inhibit tumor growth, metastasis and immune response in Lewis lung cancer mice, and its mechanism may be related to inhibiting SIRP/CD47 signaling pathway.

Biglycan as a potential diagnostic and prognostic biomarker in multiple human cancers.

Biglycan (BGN), a key member of the small leucine-rich proteoglycan family, is an important component of the extracellular matrix. Clinical studies have demonstrated that upregulation of BGN is associated with poor prognosis in patients with various types of solid cancer. The present study analyzed the mRNA expression levels of BGN in various types of solid cancer when compared with that in normal tissues via the Oncomine database. The UALCAN, OncoLnc and Kaplan-Meier Plotter databases were additionally used to evaluate the prognostic values of BGN in patients with solid cancer and co-expression gene analysis was conducted using the protein-protein interaction networks of BGN. The present study observed that the mRNA expression levels of BGN were increased in bladder, brain and central nervous system, breast, colorectal, esophageal, gastric, head and neck, lung, ovarian and 28 subtypes of cancer compared with normal tissues. The increased expression of BGN was identified to be associated with a poor outcome in ovarian and gastric cancer. Based on the co-expression network, BGN was identified as the key gene in a 43-gene network. The present findings of increased expression of BGN in solid tumors and its positive association with poor outcome on patient survival indicate that BGN may serve as a prognostic marker and as a target for novel therapeutics for multiple types of cancer.

Comparison of IDW, cokriging and ARMA for predicting spatiotemporal variability of soil salinity in a gravel-sand mulched jujube orchard.

Information about the spatiotemporal variability of soil salinity is important for managing salinization in gravel-sand mulched fields. We used inverse distance weighting (IDW) and cokriging to model the spatial variability of soil salinity from 2013 to 2016 and used an autoregressive moving-average (ARMA) model time series to analyze the temporal variability. The objectives of this paper are (a) to compare IDW and cokriging for predicting salinity in deep soil layers from surface data, thus finding a more appropriate method to model the spatial variability of soil salinity, and, using ARMA time series, (b) to identify one or a few sampling points, where soil salt content is the most temporally stable, to increase sampling efficiency or decrease cost and to estimate the overall soil salt content of a field. The IDW interpolation was more accurate than cokriging when using surface salt content to estimate the content in deep layers; so, we used IDW to interpolate the data and draw spatial distribution maps of salt content. Salinity in the 0-10 cm layer gradually decreased with the amount of gravel-sand mulching, from 1.02 to 0.7 g/kg over four years, and increased with depth. ARMA was accurate when using sample dates to predict soil salinity in the time series, and the model was more stable. The stability of the salt spatial patterns over time and along the soil profile allowed us to identify a location representative of the field-mean salt content, with mean relative error ranging between 0.56 and 2.19%. The monitoring of soil salt from a few observations is thus a valuable tool for practitioners and will aid the management of soil salt in gravel-sand-mulched fields in arid regions, with a range of potential applications beyond the framework of monitoring salinity.

Effect of various concentrations of superabsorbent polymers on soil particle-size distribution and evaporation with sand mulching.

Superabsorbent polymers (SAPs) are type of hydrogels capable to swell and absorb a large amount of water, but easily decomposed and oxidized by the air. We used electron-microscopic imaging in an indoor simulation with sand mulching to test the effects of various SAP concentrations on controlling evaporation and salt formation. The treatments were sand-mulched columns containing 0, 0.1, 0.2, 0.5 and 1.0% SAP. The soil particle pores were from dense to sparse and the corresponding fractal dimension decreased as SAP concentration increased. SAP concentration was correlated negatively with fractal dimension, clay-particle fraction and silt-volume fraction. And it showed a positive correlation with sand volume fraction. SAP concentration significantly affected the particle-size distribution. Water-storage capacity increased in each column layer (five 8-cm layers) at the same infiltration depth. Evaporation decreased the water content of each layer. Sand mulching combined with the SAP decreased evaporation in each layer relative to the control, which retained more water and decreased the accumulation of surface salt in the order 1.0% > 0.5% > 0.2% > 0.1% > 0. Salt migrated at 0-30 cm with sand mulching but 0-25 cm with sand mulching and SAP amendment. The decrease in salt accumulation was most effective at a SAP concentration of 0.2%.

A comparative study on the mechanical behavior of intervertebral disc using hyperelastic finite element model.

BACKGROUND: Lumbar disc herniation may result in excruciating pain due to it being one of the most common diseases related to changes of intervertebral disc (IVD). In order to find a better clinical treatment and prevention scheme for relieving the pain caused by spine degeneration, the mechanical behavior analysis of IVD must be studied. OBJECTIVE: A finite element method (FEM) is used in this study to analyze the mechanical behaviors of healthy and herniated IVD. METHOD: In order to compare the responses of IVD under different loading conditions for the annulus fibrosus of IVD, the hyperelastic and elastic constitutive FE models were used in the FEM. RESULTS: The comparison shows that hyperelastic FE models have a much better capability to describe the mechanical behaviors of the IVD than elastic FE models. It can be found from FE simulation that there was a higher stress concentration at the annulus fibrosus of the herniated disc than the healthy disc. CONCLUSIONS: Higher stress concentration resulted in more damage and ease of bringing out lumbar disc herniation. Numerical examples of FE simulation indicate that the FEM with hyperelastic constitutive model has very good capability for analyzing the mechanical behaviors of IVD.

Increased expression of Dock180 protein in the noninfarcted myocardium in rats.

The integrin ß1 subunit and its downstream molecule focal adhesion kinase have been identified as critical molecules for the inhibition of postinfarction cardiac remodeling, ischemic cardiomyopathy, and heart failure. However, as a component of the integrin pathway, it is still unclear whether Dock180 (dedicator of cytokinesis 1) protein is expressed in the noninfarcted myocardium of the peri-infarct zones. In this study, experimental myocardial infarction (MI) and sham-operation (sham) models were established in Sprague Dawley rats and the expression of Dock180 protein in the myocardium of the sham group and in the noninfarcted myocardium of the peri-infarct zones of the MI group was detected by Western blot technique. The Dock180 protein expression in the myocardium was as follows: postsham 24-hour group, 0.10 ± 0.04 (n = 8); post-MI 24-hour group, 0.13 ± 0.03 (n = 8); postsham 12-week group, 0.11 ± 0.05 (n = 8); and post-MI 12-week group 0.17 ± 0.04 (n = 8). The Dock180 protein expression in the myocardium in the post-MI 12-week group was significantly higher than that in the postsham 12-week group (p = 0.019), in the postsham 24-hour group (p = 0.004), and in the post-MI 24-hour group (p = 0.040). We conclude that Dock180 protein is expressed in the myocardium in rats. Furthermore, its expression is significantly increased in the noninfarcted myocardium of the peri-infarct zones.

Elevated expression of C3G protein in the peri-infarct myocardium of rats.

BACKGROUND: The integrin ß1 subunit and its downstream molecules such as integrin-linked kinase (ILK) and focal adhesion kinase (FAK) are indispensable to the inhibition of postinfarction cardiac remodeling, ischemic cardiomyopathy, and heart failure. As a component of the integrin pathway, C3G (Crk SH3-domain-binding guanine nucleotide exchange factor) protein may also participate in postinfarction cardiac remodeling, ischemic cardiomyopathy, and heart failure. MATERIAL/METHODS: Experimental myocardial infarction (MI) and sham-operation (sham) models were set up in Sprague-Dawley rats. C3G protein expression in the myocardium in the sham group and in the non-infarcted myocardium of the peri-infarct zones in the MI group was examined by Western blot. RESULTS: The C3G protein expression in the myocardium was 0.22±0.06, n=8 in the post-sham 24-hour group; 0.29±0.10, n=8 in the post-MI 24-hour group; 0.22±0.07, n=8 in the post-sham 12-week group; and 0.56±0.14, n=8 in the post-MI 12-week group. The C3G protein expression in the myocardium in the post-MI 12-week group was significantly elevated compared to that in the post-sham 12-week group (p=0.0002), in the post-sham 24-hour group (p=0.0002), and in the post-MI 24-hour group (p=0.0006). CONCLUSIONS: C3G protein expression exhibits in the myocardium of rats. Furthermore, C3G protein expression is significantly elevated in the non-infarcted myocardium of the peri-infarct zones. The elevated C3G protein expression could participate in postinfarction cardiac remodeling, ischemic cardiomyopathy, and heart failure.

Sintering photonic beads for multiplex biosensing.

Monodispersed silica nanoparticles were assembled into opaline photonic crystal beads and cemented together by sintering in order to enhance the mechanical stability of the beads, which simulates the formation of nature opal. The sintering temperatures and structures of the sintered beads were investigated. The results showed that by sintering treatment at temperatures between 700 degrees C and 1000 degrees C for 2 hours the beads were more reliable for multiplex bioassay applications in mechanical robustness, fluorescent background, sensitivity and encoding ability. It is anticipated that the opal-like beads have wide applications in multiplex protein and nucleic acid analysis.

Multiplex detection of tumor markers with photonic suspension array.

A novel photonic suspension array was developed for multiplex immunoassay. The carries of this array were silica colloidal crystal beads (SCCBs). The codes of these carriers are the characteristic reflection peak originated from their structural periodicity, and therefore they do not suffer from fading, bleaching, quenching, and chemical instability. In addition, because no dyes or materials related with fluorescence are included, the fluorescence background of SCCBs is very low. With a sandwich format, the proposed suspension array was used for simultaneous multiplex detection of tumor markers in one test tube. The results showed that the four tumor markers, alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), carcinoma antigen 125 (CA 125) and carcinoma antigen 19-9 (CA 19-9) could be assayed in the ranges of 1.0-500 ng mL(-1), 1.0-500 ng mL(-1), 1.0-500 U mL(-1) and 3.0-500 U mL(-1) with limits of detection of 0.68 ng mL(-1), 0.95 ng mL(-1), 0.99 U mL(-1) and 2.30 U mL(-1) at 3 sigma, respectively. The proposed array showed acceptable accuracy, detection reproducibility, storage stability and the results obtained were in acceptable agreement with those from parallel single-analyte test of practical clinical sera. This technique provides a new strategy for low cost, automated, and simultaneous multiplex immunoassay.