Collagen is the main constituent of gelatinous Chinese medicine, with deer hide gelatin (Cervi Corii Colla, DHG) made from deer hide (DH) through a complex thermal and high-pressure processing procedure. During this procedure some amino acids in collagen undergo hydroxylation and deamidation. In the present study, comparative analysis of proteins and peptides in DH and DHG was carried out using "peptidomics-modifications" methods. Nano-LC-MS/MS was used to analyze proteins and peptides in DH and DHG, and the number and sites of modification were determined as well. The amount of hydroxylation and deamidation that occurred in DHG was significantly greater than that in DH, suggesting that under thermal and high-pressure processing these modifications occurred more frequently on certain amino acids in collagen, and might be correlated with hydrophobicity. The occurrence and mechanism of hydroxylation and deamidation in DH processing procedures should be explored in further research. The present study provides important evidence of the chemical constituents and the correlation of processing procedures with these modifications, and also suggests some investigative ideas for DHG processing optimization and improvement of quality standards.

Ethylene-response factors, which are a subfamily of the AP2/ERF family, play an important role in ethylene signal transduction, plant growth and plant resistant. In this study, a full-length cDNA of the AsERF1 gene was cloned from Aquilaria sinensis. Sequence analysis, prokaryotic expression and purification, subcellular localization, tissue-specific analysis and expression analysis under different abiotic stresses was performed. The open reading frame (ORF) of the AsERF1 gene was 691 bp, encoding a protein of 229 amino acids with a predicted molecular mass of 25.36 kD. The AsERF1 protein contained the conserved AP2 sequence of ERF protein. A phylogenetic analysis indicated that the AsERF1 protein showed greatest sequence similarity with ERF2 from Populus trichocarpa. The recombinant AsERF1 protein was expressed in Escherichia coli BL21(DE3) cells using the prokaryotic expression vector pET28a-AsERF1 and the recombinant AsERF1 protein was purified. Agrobacterium-mediated protein expression experiments demonstrated that AsERF1 mainly localized to the nucleus. Expression analysis indicated that AsERF1 was primarily observed in leaves. The AsERF1 expression level was induced by salt, drought, low temperature and CdCl₂ treatment, while the abundance of AsERF1 was most significantly induced by drought stress. These results provide valuable insights into the role of AsERF1 in plant defense and the mechanism of agarwood formation.

Ginsenoside Ro decreased measures of inflammation, aging, oxidants and thrombus formation in a previous study. To measure ginsenoside Ro content in red ginseng from different years, an optimized extraction method was developed to determine ginsenoside Rg₁, Re, Rb₁ and Ro content by HPLC in 43 batches of red ginseng from different origins, growing years and manufacturers. The results indicate that the best extraction method was to ultrasonify a 1 g sample in 70% methanol for 50 min. The total running time of the optimized gradient was 50 min using a C18 core-shell column and was half the time described in the Chinese Pharmacopoeia, 2015 edition. The separation resolution of all of targeted compounds was greater than 1.6. The peak shape of ginsenoside Ro was optimal when the mobile phase consisted of acetonitrile and water with 0.1% phosphoric acid. The content of ginsenoside Ro was in the range of 0.11% to 0.43%, and the average content was 0.26%, which was higher than that of ginsenoside Rg₁ and Re. The ratio of ginsenoside Ro and Re as a threshold could be used to discriminate red ginseng from different growing years; in addition, 100%, 94.4% and 46.6% of red ginseng from six, five and four years exceeded the threshold of 1.3. Our optimized analytical method for characterization of red ginseng is convenient and shortens the assay time.

Drug transporters and metabolic enzymes are the key proteins in the disposition of drugs in the body. In recent years it has been found that there is a cooperative relationship between drug transporters and metabolizing enzymes. Functional changes in drug transporters or metabolizing enzyme can affect the ability to eliminate drugs. Therefore, it is important to clarify this cooperative relationship, which is directly related to the pharmacokinetics, pharmacodynamics and adverse effects of drugs. Intestine and liver are the main organs of drug metabolism. There are abundant drug transporters and metabolizing enzymes in the tissues. This paper reviews the influence of the cooperative relationship between drug transporters and metabolizing enzymes on drug disposition by intestine and liver.

In recent years, multi-modal combined anti-tumor has become an effective strategy for clinical tumor treatment. Photothermal therapy with its characteristics of minimally invasive, controllable, high efficiency, and strong specificity, can effectively make up for the toxic side effects and tumor resistance caused by traditional drug treatment. The research shows that the combination of photothermal therapy and chemotherapy has better synergistic antitumor effect. However, chemotherapeutic drugs and photothermal agents may have different pharmacokinetic behaviors in vivo, so it is difficult to ensure their effective transmission in tumor site, and the free form is easy to be metabolized and degraded in vivo. How to deliver the two therapeutic modes of drugs/photothermal reagents to tumor tissues in a specific, efficient and synchronous manner to achieve the best combined antitumor effect is an important problem to be solved in the combined antitumor application. The development of nano-drug delivery technology provides a new idea for the application of tumor treatment. In this paper, combined with the latest research progress in this field, the anti-tumor mechanism of photothermal therapy combined with chemotherapy, the advantages of nano drug delivery, the types and characteristics of commonly used nano materials and the principle of drug delivery are reviewed in order to provide a reference for the further development of multi-mode combined treatment of tumor.

To detect the methylation level of genome-wide DNA and total RNA in the process of heart failure, we established the method of ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) to observe the change and synchronization of methylation rate of myocardial infarction (MI) tissue and peripheral blood. Animal welfare and experimental process were in accordance with the regulations of the Animal Ethics Committee of Guangzhou Medical University. The rats with myocardial infarction were divided into three groups:1^st, 4^th, and 8^th week to simulate different levels of cardiac function. And they were euthanized at the same time to keep the same age. DNA and RNA were extracted from infarct marginal tissues and peripheral blood lymphocytes, and then decomposed into single nucleosides by enzymolysis. The methylation rate of DNA and RNA was measured and calculated quantitatively. The results showed a concordant methylation changes in tissue and blood, and the methylation level of genome-wide DNA and total RNA was increased after myocardial infarction in rats. In this study, we obtained the preliminary data of DNA and RNA methylation during the occurrence and development of heart failure, further indicating that epigenetic changes can be used as biomarkers for early diagnosis of heart failure.

To establish a method for the determination of polymer impurities in ceftazidime raw materials and preparations, a ceftazidime degradation solution containing polymer impurities was prepared by forced polymerization. Polymer impurities in the degradation solution were separated and identified by high performance gel chromatography and the column switching-LC-MSⁿ method. A new RP-HPLC method for ceftazidime polymer was established and validated with a Phenomenex Gemini-C18 column using a mobile phase gradient elution of 0.02 mol·L^-1 phosphate buffer, methanol and acetonitrile. The results showed that when using this high performance gel chromatography method some small molecular weight impurities were co-eluted with the polymers, resulting in a poor specificity and poor quantitative accuracy. But when using the RP-HPLC method, four polymer impurities were detected in the 25-45 min time range with good specificity, sensitivity and robustness, including two ceftazidime dimers, trimers, and derivatives. Therefore, the described RP-HPLC method is suitable for the quality control of polymer impurities in ceftazidime, and ceftazidime degradation solution can be used as suitable solution for analysis of ceftazidime polymers.

N-Acetylaspartate (NAA) is a highly abundant brain metabolite. Nowadays, as an important marker reflecting the function of nervous system, NAA is widely used in the results analysis of nuclear magnetic resonance spectroscopy (¹H MRS). NAA is synthesized in mitochondria of neurons and metabolized in oligodendrocytes. Additionally, NAA may be converted to the dipeptide N-acetylaspartylglutamate (NAAG), and catabolized into NAA and glutamate in astrocytes. NAA is related to a variety of central nervous system diseases, including Canavan disease, multiple sclerosis, depression, schizophrenia and other mental diseases. Therefore, NAA may be a biomarker of these diseases, and its related enzymes may be used as therapeutic targets for drug screening. Here, we combined the current research on the molecular mechanisms of NAA to reveal the process of NAA generation, metabolism and transport in the brain, explain the possible physiological effects of NAA and discuss its relationship with central nervous system diseases, explore the prospect of NAA in disease prediction and diagnosis, as well as the targeted treatment that may become the breakthrough of refractory diseases.

Animal derived traditional Chinese medicines (ATCMs) are an important part of traditional Chinese medicine (TCM). The lack of proper ideas and strategies made it not systematic and perfect enough on investigating bioactive components and quality evaluation of ATCMs, which restrict many aspects of ATCMs investigation including clinical applications, pharmaceutical technologies, and quality control. Therefore, based on our previous investigations of animal horn and animal derived gelatin TCMs, and the research progress at home and abroad, ideas and strategies for investigating the correlations between proteins/peptides and their bioactivities in animal horn and animal gelatin derived TCMs based on integrated "proteomics/peptidomics-modifications" methods was proposed. Firstly, proteomics and peptidomics analysis can be used to study proteins and peptides in ATCMs. Secondly, modification analysis can be used to reveal those chemical modifications on proteins and peptides of ATCMs. Thirdly, the correlations between components, modifications and traditional bioactivities can be systemic discussed. Based on the present study, hopefully, enough evidences and reference can be provided to resolve the issues in ATCMs investigations on modernization and bioactive material basis.

We observed the effect of calcium dobesilate (CaD) on apoptosis induced by cisplatin in human proximal tubular epithelial cells (HK-2) and explored the possible mechanism. Based on HK-2 cells apoptosis model induced by cisplatin, CCK-8 method was used to detect the effect of CaD on the proliferation of HK-2. Apoptosis was detected by flow cytometry. Reactive oxygen species (ROS) assay was used to evaluate the level of oxidative stress. The mitochondrial membrane potential was measured by JC-1 method. The expression levels of p53, caspase-3, bcl-2 and bax in cisplatin-induced HK-2 were detected by Western blot. The expression of renal injury factor 1(KIM-1) and neutrophil gelatin-related apolipoprotein (NGAL), markers of acute kidney injury, were detected by ELISA. The results showed that CaD could reduce the oxidative stress level induced by cisplatin and inhibit apoptosis in renal tubular epithelial cells. Cisplatin can up-regulate the protein expressions of p53, caspase-3, bax, KIM-1 and NGAL, and reduce the expression of bcl-2. After using CaD, the protein levels of KIM-1, NGAL, p53, caspase-3 and bax were significantly reduced, while the levels of bcl-2 were increased. This study has shown that CaD can alleviate cisplatin-induced HK-2 injury and inhibit HK-2 apoptosis, which may be related to the regulation of bax/bcl-2/caspase-3 apoptosis signaling pathway.