Characterization of a gene encoding clathrin heavy chain in maize up-regulated by salicylic acid, abscisic acid and high boron supply.

Clathrin, a three-legged triskelion composed of three clathrin heavy chains (CHCs) and three light chains (CLCs), plays a critical role in clathrin-mediated endocytosis (CME) in eukaryotic cells. In this study, the genes ZmCHC1 and ZmCHC2 encoding clathrin heavy chain in maize were cloned and characterized for the first time in monocots. ZmCHC1 encodes a 1693-amino acid-protein including 29 exons and 28 introns, and ZmCHC2 encodes a 1746-amino acid-protein including 28 exons and 27 introns. The high similarities of gene structure, protein sequences and 3D models among ZmCHC1, and Arabidopsis AtCHC1 and AtCHC2 suggest their similar functions in CME. ZmCHC1 gene is predominantly expressed in maize roots instead of ubiquitous expression of ZmCHC2. Consistent with a typical predicted salicylic acid (SA)-responsive element and four predicted ABA-responsive elements (ABREs) in the promoter sequence of ZmCHC1, the expression of ZmCHC1 instead of ZmCHC2 in maize roots is significantly up-regulated by SA or ABA, suggesting that ZmCHC1 gene may be involved in the SA signaling pathway in maize defense responses. The expressions of ZmCHC1 and ZmCHC2 genes in maize are down-regulated by azide or cold treatment, further revealing the energy requirement of CME and suggesting that CME in plants is sensitive to low temperatures.

Label-free SERS strategy for rapid detection of capsaicin for identification of waste oils.

Identification of waste oils is challenging in the field of food safety due to the lack of common markers and straightforward analytical methods. Herein, we developed a novel label-free surface-enhanced Raman spectroscopy (SERS) strategy to identify waste oils using Ag nanoparticles solution (Ag NPs sol.) as a SERS substrate to significantly enhance the Raman signal of capsaicin marker molecule usually contained in the waste oils. The enhanced signal was directly detected by a portable Raman spectrometer with the limit of detection (LOD) of 2.9 µg L-1 within 10 min. Concentration-dependent SERS investigation showed the linear relationship between the SERS signal intensity of the characteristic peaks and the concentrations of capsaicin in the range of 10-2500 µg L-1 and the correlation coefficient was 0.9895. Our findings show the sensitivity, accessibility, and reliability of this method for the rapid identification of waste oils and furthermore for the practical applications in the field of food safety.

Rapid and Quantitative Detection of Aflatoxin B1 in Grain by Portable Raman Spectrometer.

Many foodstuffs are extremely susceptible to contamination with aflatoxins, in which aflatoxin B1 is highly toxic and carcinogenic. Therefore, it is crucial to develop a rapid and effective analytical method for detecting and monitoring aflatoxin B1 in food. Herein, a surface-enhanced Raman spectroscopic (SERS) method combined with QuEChERS (quick, easy, cheap-effective, rugged, safe) sample pretreatment technique was used to detect aflatoxin B1. Sample preparation was optimized into a one-step extraction method using an Au nanoparticle-based solution (Au sol) as the SERS detection substrate. An affordable portable Raman spectrometer was then used for rapid, label-free, quantitative detection of aflatoxin B1 levels in foodstuffs. This method showed a good linear log relationship between the Raman signal intensity of aflatoxin B1 in the 1-1000 µg L-1 concentration range with a limit of detection of 0.85 µg kg-1 and a correlation coefficient of 0.9836. Rapid aflatoxin B1 detection times of ∼10 min for wheat, corn, and protein feed powder samples were also achieved. This method has high sensitivity, strong specificity, excellent stability, is simple to use, economical, and is suitable for on-site detection, with good prospects for practical application in the field of food safety.

Protective effect of reduced glutathione C60 derivative against hydrogen peroxide-induced apoptosis in HEK 293T cells.

Hydrogen peroxide (H2O2) and free radicals cause oxidative stress, which induces cellular injuries, metabolic dysfunction, and even cell death in various clinical abnormalities. Fullerene (C60) is critical for scavenging oxygen free radicals originated from cell metabolism, and reduced glutathione (GSH) is another important endogenous antioxidant. In this study, a novel water-soluble reduced glutathione fullerene derivative (C60-GSH) was successfully synthesized, and its beneficial roles in protecting against H2O2-induced oxidative stress and apoptosis in cultured HEK 293T cells were investigated. Fourier Transform infrared spectroscopy and (1)H nuclear magnetic resonance were used to confirm the chemical structure of C60-GSH. Our results demonstrated that C60-GSH prevented the reactive oxygen species (ROS)-mediated cell damage. Additionally, C60-GSH pretreatment significantly attenuated H2O2-induced superoxide dismutase (SOD) consumption and malondialdehyde (MDA) elevation. Furthermore, C60-GSH inhibited intracellular calcium mobilization, and subsequent cell apoptosis via bcl-2/bax-caspase-3 signaling pathway induced by H2O2 stimulation in HEK 293T cells. Importantly, these protective effects of C60-GSH were superior to those of GSH. In conclusion, these results suggested that C60-GSH has potential to protect against H2O2-induced cell apoptosis by scavenging free radicals and maintaining intracellular calcium homeostasis without evident toxicity.

Expression of IGF-II in early experimental hepatocellular carcinomas and its significance in early diagnosis.

AIM: To investigate the serum level and expression of insulin growth factor II (IGF-II) in liver tissues of rats with early experimental hepatocellular carcinomas (HCC) and its significance in early diagnosis. METHODS: Early experimental hepatocellular carcinomas were induced by diethylnitrosamine (DENA) in 180 male SD rats. Another 20 male SD rats served as control. The IGF-II serum level was measured by ELISA. Immunohistochemistry and electron microscopic immunohistochemistry were used to observe the expression of IGF-II in normal and tumor liver tissues and its ultrastructural location in malignant hepatocytes. The expressions of IGF-II in human hepatoma cell lines HepG2, SMMC7721 and human embryonic liver cell line L-02 were measured by immunocytochemistry. IGF-II mRNA level was studied by in situ hybridization. RESULTS: IGF-II was expressed in the cytoplasm of both sinusoidal cells in paracancerous cirrhotic liver tissue and malignant hepatocytes in early experimental HCC tissues. Gold particles were seen on the rough endoplasmic reticulum and the mitochondrion in malignant hepatocytes. IGF-II was expressed in the human hepatoma cell lines. The mRNA level of IGF-II was higher in rat liver tumor tissue than in normal rat liver tissue. The serum IGF-II level of the early experimental HCC group was 34.67+/-10.53 ng.ml(-1) and that of the control group was 11.75+/-5.84 ng.ml(-1). The rank sum test was used for statistical analysis. There was a significant difference between the two groups (P<0.01). CONCLUSION: During the induction of early experimental HCC by DENA, IGF-II may promote hepatocytic proliferation via a paracrine mechanism in the pre-cancerous stage. When hepatocytes are transformed into malignant cells, they may secrete IGF-II and promote malignant cell proliferation by an autocrine mechanism. IGF-II may be a possible biological marker in the early diagnosis of HCC.

[Effect of electroacupuncture pretreatment on glutamate-NMDAR signal pathway in hippocampal neurons of vascular dementia rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) pretreatment on hippocampal glutamate (Glu) content and N-methyl-D-aspartate receptor (NMDAR)1 mRNA expression in rats with vascular dementia (VD) so as to explore its underlying mechanism in improving VD. METHODS: A total of 72 SD rats were randomized into control (n=16), sham-operation (n=16), model (n=20) and EA pretreatment (n=20) groups. VD model was established by modified middle cerebral artery occlusion. EA (1 mA, 1.7 Hz) was applied to "Baihui" (GV 20), "Shenshu"(BL 23) and "Zusanli" (ST 36), once daily for 10 days. Glu content of right hippocampus tissue was detected with chromatometry. NMDAR 1 mRNA expression of hippocampus was detected by in situ hybridization histochemistry. RESULTS: In comparison with sham operation group, Glu content and NMDAR 1 mRNA expression of hippocampus in model group increased significantly (P<0.01). Compared with model group, Glu content and NMDAR 1 mRNA expression of hippocampus in EA pretreatment group decreased significantly (P<0.01, 0.05). CONCLUSION: EA pretreatment can suppress the increase of Glu content, down-regulate NMDAR 1 mRNA expression in VD rats, which may contribute to its effect in relieving VD via reducing apoptosis and protecting cerebral neurons.