ABSTRACT
This paper aims to systematically analyze the peptides and proteins from Asini Corii Colla(ACC) through shotgun proteomics. After high-pH reversed-phase fractionation, the proteins and peptides in the hydrolysate of ACC were further separated by nano LC-Q-Exactive-MS/MS under the following conditions: Thermo Scientific EASY column(100 μm×2 cm, 5 μm, C_(18)) as precolumn, Thermo Scientific EASY column(75 μm×100 mm, 3 μm, C_(18)) for solid phase extraction, gradient elution with 0.1% formic acid in water(mobile phase A) and 84% acetonitrile in water containing 0.1% formic acid(mobile phase B), and MS in positive ion mode. Based on Uniprot_Equus caballus, MS data, and literature, 2 291 peptides were identified from ACC by MaxQuant, with 255 Maillard reactions(AML, CML, CEL)-modified peptides identified for the first time. Through alignment, the peptides were found to belong to 678 equine proteins. In conclusion, the combination of nano LC-Q-Exactive-MS/MS and shotgun proteomics achieved rapid and accurate identification of the proteins and peptides in ACC, which provides the key information and new insights for further investigation of chemicals and effective substances in ACC.
Subject(s)
Animals , Chromatography, Liquid , Horses , Peptides , Proteins , Proteomics , Tandem Mass SpectrometryABSTRACT
Objective To explore the nervous regulating mechanism in the effect of light intensity on reproductive performance. Methods Sixty healthy Jinghong-1 hens aged 100 days were randomly divided into four groups of 15 hens and applied a light of 30 lx, 20 lx, 10 lx, and 1 lx, respectively, for 7 days. The distribution and expression of gonadotropin-releasing hormone (GnKH) in mesencephalon and diencephalon were detected using immunohistochemical and image analysis methods. Results GnKH immunoreactive positive neurons were mainly observed in stratum griseum centrale (SCC) of tectum, nucleus isthmi pars magnocellularis (Imc) , nucleus isthmi pars parvocellularis (Ipc) , nucleus subpre-tectalis/interstito-pretectalis-subpretectalis (SP/IPS), nucleus of basal optic root (nbor), and nucleus semilunaris (Slu) of mesencephalon. The mean gray values of positive neurons in 30 lx and 20 lx groups were significantly higher than those of 10 lx and 1 lx groups (P0. 05) between 30 lx and 20 lx groups, but they were significant higher than those of 10 lx and 1 lx (P<0. 05) in PHN, PVN, and GLv. The percent cell area in Hot was the largest in 30 lx group, having no significant difference comparing 20 Ix, being significant higher than those of 10 lx and I Ix (P<0. 05). Those in 30 lx and 20 lx groups were significant higher than 10 lx and 1 lx (P<0.05) in PHN and PVN. Conclusion Light intensity affect the expression of GnRH in hen mesencephalon and diencephalon obviously in Jinghong-1 hens aged 100 days, regulating their reproductive function. The tectofugal pathway of the optical pathways, including collateral circulation of nucleus isthmi, plays an important role in the course of regulation.
ABSTRACT
Traditional Chinese medicine( TCM) glues,including leather glues,horn glues,nail glues and bone glues,have a long application history and unique characteristics. In recent years,their market demand has increased year by year because of their remarkable curative efficacy and nourishing effects,which leads to insufficient supply of raw material resources,and widespread use of fake and inferior products,seriously affecting the reputation of TCM glues and drug safety. In this context,the establishment of a more specific quality detection method for the TCM glues according to their specific characteristics can effectively improve the quality control level,promote rational use,and have a far-reaching impact on the industrial development of TCM glues. In this paper,the classification of TCM glues,as well as the production and application status of their representative( Ejiao) were briefly introduced; the papers on quality control technologies of TCM glues,including traditional identification experience,authenticity identification,physical property determination,protein,peptide and amino acid contents determination,element analysis,biological evaluation,and brand protection technology of TCM glues,were reviewed,and their advantages and disadvantages were summarized and analyzed comprehensively.Based on the specific characteristics of TCM glues,such as complex material basis,unclear pharmacodynamic components and different production processes,it was proposed in this paper to research and develop information-rich,convenient,fast,and non-destructive analytical techniques for the quality control of TCM glues and brand protection of famous products,thus promoting the healthy development of TCM glues industry.
Subject(s)
Adhesives , Medicine, Chinese Traditional , Quality Control , ResearchABSTRACT
<p><b>OBJECTIVE</b>To observe the biological role and underlying mechanism of microRNA-132 (miR-132) in liver cancer cell proliferation and apoptosis.</p><p><b>METHODS</b>The expressions of miR-132 in the cancer tissue and their adjacent tissues from 45 liver cancer patients were detected by real-time quantitative polymerase chain reaction (RT-qPCR). The biological effects of miR-132 transfection on human liver cancer MHCC97H cells were assessed by CCK-8 assay, flow cytometry,in vivo experiment in nude mice, and TUNEL test. Western blotting was used to detect the expressions of p-AKT, Survivin, and Caspase 3 in liver cancer cells. Immunohistochemistry was used to detect the positive expressions of Ki-67,Survivin,and Caspase 3 in the xenograft tumors.</p><p><b>RESULTS</b>The expression level of miR-132 was found to be down-regulated in liver cancer tissues compared with the matched adjacent tissues (P<0.05). After transfection,the expression of miR-132 was significantly higher than blank control group and negative control group (P<0.05). The proliferation of liver cancer cells was inhibited significantly by miR-132 transfection (P<0.05). Transfection of miR-132 arrested cells in the G0/G1 phase and triggered apoptosis of MHCC97H cells (P<0.05). After miR-132 transfection,the expression of Caspase 3 was up-regulated, whereas the expressions of p-AKT and Survivin were down-regulated (P<0.05). In addition,the tumor weight in miR-132 transfection group was significantly decreased in comparison with blank control group and negative control group (P<0.05). Apoptosis occurred more frequently in the miR-132 transfection group than in control groups (P<0.05). Compared with the blank control group and negative control group, the miR-132 transfection group had significantly decreased expression of Survivin but increased positive expression of Ki-67 and Caspase 3(P<0.05).</p><p><b>CONCLUSIONS</b>miR-132 is down-regulated in human liver cancer tissues miR-132 transfection can effectively inhibit proliferation and promote apoptosis of MHCC97H cells in vitro and in vivo. Therefore, miR-132 may become a new target in liver cancer treatment.</p>