Two new disubstituted maleimides, aspergteroids G–H (1–2), and two trisubstituted butenolides aspergteroids I–J (3–4), along with four known analogs, were isolated and structurally identified from the fermentation extract of soft-coral-associated symbiotic and epiphytic fungus Aspergillus terreus EGF7-0-1. The structures of the new compounds were established mainly via spectroscopic data analyses, and their absolute configurations were determined via X-ray diffraction analysis and comparison of the calculated and experimental electronic circular dichroism. Myocardial protection assays showed that compounds 1, 2, 5, and 6 possess protective effects against tert-butyl hydroperoxide (TBHP)-induced H9c2 (rat myocardial cells) apoptosis at low concentrations. Based on the analyses of the protein–protein interaction (PPI) network and Western blotting, compound 1 may inhibit the apoptosis and inflammatory response of cardiomyocytes after TBHP induction and improve the antioxidant capacity of cardiomyocytes. We speculate that the anti-inflammatory response of compound 1 is suppressed by the glycogen synthase kinase-3 beta (GSK-3β), downregulated by the NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome activation, and suppressed by the expression of cysteinyl aspartate specific proteinase-3 (caspase-3) and B-cell lymphoma-2 associated X protein (Bax).

Chinese herbal medicines (CHMs) play an important role in the clinical efficacy of TCM. It is necessary to establish an intelligent identification method for CHMs to assist in Chinese medicine dispensing. In the present study, 160 kinds of CHMs were collected from the real world, and an image database containing 44467 pictures was constructed. Based on the L18 (3^6,6^1) hybrid orthogonal table, an orthogonal experiment was carried out on seven influencing factors, namely architecture, learning rate, optimizer, weight decay, batch size, gradual unfreezing, and discriminative fine-tuning. Five-fold cross-validation was used to calculate the accuracy rate, recall rate, and other evaluation indicators, and in this way, the optimal model and hyperparameter combination were screened. It was found that the optimal model was ResNeXt-152. When the batch size was 64, the learning rate was 0.00005, weight decay was 0.0005, gradual unfreezing was 7, discriminative fine-tuning was 3, and optimizer was AdaMax, which was the optimal combination of hyperparameters. At this time, the highest accuracy rate was 95.36%. This shows that deep learning model training based on orthogonal design can reduce the number of experiments while improving classification accuracy.

INTRODUCTION:The fundamental theory of traditional Chinese medicine (TCM) implies that when different diseases have the same pathogen, the syndromes of these individual diseases will be the same. "Treating different diseases with the same method" is a TCM principle suggesting that when different diseases have similar pathological changes during different stages of their development, the same method of treatment can be applied. Our study aims to analyze the concept "treating different diseases with the same method" from a molecular perspective, in order to clarify its biological basis and to objectively standardize future TCM syndrome research.OBJECTIVE:The TCM syndromes Qi deficiency and blood stasis have similar pathogenesis in relation to coronary heart disease (CHD) and stroke. We aim to use big data technology and complex network theory to mine the genes specifically relevant to these TCM syndromes. This study aims to explore the correlation between the biological indicators of CHD and stroke from a scientific perspective.METHODS:Mining the relevant neuroendocrine-immune (NEI) genes by means of gene entity recognition, complex network construction, network integration, and decomposition to categorize relevant syndrome terms and establish a digital dictionary of gene specifically related to individual diseases. We analyzed the biological basis of "treating different diseases with the same method" from a molecular level using the TCMIP v2.0 platform in order to categorize the TCM syndromes most relevant to CHD and stroke.RESULTS:We found 46 genes were involved in the TCM syndromes of Qi deficiency and blood stasis of CHD and stroke. The same genes and their molecular mechanism also appeared to be in close relation to inflammatory response, apoptosis, and proliferation.CONCLUSION:By using information extraction and complex network technology, we discovered the biological indicators of the TCM syndromes Qi deficiency and blood stasis of CHD and stroke. In the era of big data, our results can provide a new method for the researchers of TCM syndrome differentiation, as well as an effective and specific methodology for standardization of TCM.

Objective: To explore the effects of Peiyuan Tongnao (PYTN) Capsule on working memory, the content of glutamic acid, and the expression of NMDA receptor 2B (NR2B) in rats with cerebral ischemia. Materials and Methods: 52 Sprague-Dawley rats were randomly divided into sham group, model group, nimodipine group, and PYTN group. The bilateral common carotid artery occlusion (BCAO) was performed to establish rat model with cerebral ischemia in the model, nimodipine, and PYTN groups. Gastric lavage with some drug based on body weight conversion was performed daily for 4 weeks in the nimodipine and PYTN groups. The working memory of the rats was tested by Morris water maze. The expression of Nissl body in hippocampus tissue was observed by Nissl staining. The determination of Glu content in hippocampus was detected by high-performance liquid chromatography. The expression level of NR2B in hippocampus area was determined using Western blot. Results: Morris water maze test on working memory escape latency. In the model group, day 1 versus day 4, there was a statistical difference (P < 0.05). In the nimodipine group, day 1 vs. day 4 displayed a statistical difference (P < 0.05). In the PYTN group, day 1 versus day 3 and day 1 versus day 4 were in significant differences (P < 0.05). The Glu content in hippocampus of the sham group was significantly different from that of model group (P < 0.05). The Glu content in the PYTN group was significantly different from that of the model group (P < 0.05). With regard to the expression of NMDA 2B in hippocampus between the sham group and the model, nimodipine, and PYTN groups, all were displayed statistical significance (P < 0.01). As the same, the expression of NMDA 2B in the model group was significant from that of the nimodipine group (P < 0.05) and PYTN group (P < 0.01). Conclusion: PYTN capsule was beneficial for improving working memory and protect neural cells in rats of cerebral ischemia, which may be associated with upregulation of the expression of Glu and NMDAR2B in hippocampus.

One-year-old Glycyrrhiza uralensis Fisch, ex DC, Fabaceae. was treated with three exogenous phytohor-mones in June and July, namely gibberellin. auxin (indole-3-acetic acid), methyl jasmonate at different concentrations. Control plants were treated with water. Roots of controls and hormones-treated C. uralensis plants were harvested at different times, and the contents of seven main chemical components were determined. Root glycyrrhizic acid content of plants treated in June increased significantly compared with controls, and the difference was significant. As for plants treated in July, root glycyrrhizic acid content increased in which sprayed with appropriate concentrations of hormones, but the effects of hormones were more evident in plants treated in June coincided with the vigorous growth period than those treated in July. Gibberellin at 40 mg/l and auxin at 40 mg/l applied in the two treatment periods significantly promoted the accumulation of glycyrrhizic acid in C. uralensis root. Treatment with methyl jasmonate at 100 and 25 mg/l in June and July, respectively, also increased glycyrrhizic acid content significantly. The determination of major active compositions indicated that liquiritin. isoliquiritin. isoliquiritin apioside and liquiritin apioside contents were positively related to glycyrrhizic acid content. The study preliminarily found phytohormones and the main chemical components associated with glycyrrhizic acid content, and these discoveries could provide a basis for establishing a chemical control network with glycyrrhizic acid as the core, confirming the secondary product metabolic pathways in the network and completely uncovering synthesis mechanism underlying glycyrrhizic acid-combined functional gene polymorphism.

Objective: Oxidative DNA damage is involved in mutation, cell death, carcinogenesis, and aging. Since biflavonoid amentoflavone exhibited beneficial effects on DNA, we therefore investigated its protection against center dot OH-induced DNA oxidative damage then discussed the mechanism. It will enhance the understanding of interaction between biflavonoid vs DNA mediated by free radicals.Methods: The protective effect of amentoflavone against center dot OH-induced DNA damage was measured using our method. To explore the mechanism, it was further determined by center dot OH-induced bases damage, deoxyribose damage, and various antioxidant assays.Results: Amentoflavone increased dose-dependently its protective percentages against center dot OH-induced damage on DNA, bases, and deoxyribose; The IC50 were 31.85 +/- 4.75, 198.75 +/- 33.53, 147.14 +/- 20.95, 75.15 +/- 10.52, 93.75 +/- 16.36, 167.69 +/- 13.90, and 137.95 +/- 19.86 mu M, respectively for DNA, cytosine, uracil, adenine, thymine, guanine, and deoxyribose damages. Radical-scavenging assays suggested that amentoflavone could effectively scavenge center dot O-2(-), DPPH center dot, ABTS center dot(+) radicals (IC50 values were respectively 8.98 +/- 0.23, 432.25 +/- 84.05, 7.25 +/- 0.35 mu M).Conclusions: Based on the mechanistic analysis, it is concluded that amentoflavone can effectively protect against center dot OH-induced oxidative damage DNA (including base & deoxyribose moieties), via deoxynucleotide radical repairing, and reactive oxygen species (ROS) scavenging approaches which may be mediated by donating hydrogen atom (H center dot) and electron (e). Further analysis indicated that both scavenging and repairing approaches can be primarily attributed to its antioxidant mechanism which may ultimately arise from to the stability of its oxidized product semi-quinone form. Its protection against DNA damage may be generally responsible for the radioprotective and anti-inflammation effects.

Previous deoxyribose degradation method lacks biological relevance, specificity, and even reliability. In this study, a new hydroxyl radical-scavenging assay based on DNA damage is described. 2-Thiobarbituric acid-reactive substance (TBARS, λmax = 530 nm) was chosen as the biomarker of hydroxyl-mediated DNA damage. On the basis of systematic investigations into various factors affecting A530 nm and solvent interference, the experimental procedure was developed. The successful measurement of 30 selected antioxidants demonstrated that the proposed DNA damage method is reliable, simple, specific, and biologically relevant. It is suitable for all types of antioxidants in vitro.

(2 S,2″ S)-Tetrahydroamentoflavone (THA) is a typical biflavonoid. In this study, we systematically evaluated its antioxidant activity by various antioxidant assays in vitro. The IC 50 values of THA were 4.8±0.3, 743.2±49.5, 35.5±1.9, 165.7±22.8, 4.4±0.2, and 77.1±2.2 μg/mL, respectively, for superoxide (•O 2 - ) radical-scavenging, Fe 2+ -chelating, Cu 2+ -chelating, DPPH• (1,1-diphenyl-2-picrylhydrazyl radical) scavenging, ABTS• + (3-ethylbenzthiazoline-6-sulfonic acid radical) scavenging, and Cu 2+ -reducing power. The average ratio value of IC 50, Trolox :IC 50, THA was calculated as 2.54, suggesting that THA possesses 2.54 times the total antioxidant level than the standard antioxidant Trolox. THA exerts its antioxidant activity in vitro through metal-chelating, and radical-scavenging, which is via donating a hydrogen atom (H•) and an electron (e). Its antioxidant activity can be responsible for its pharmacological effect and make it an attractive natural antioxidant.

Rhizoma Atractylodes macrocephala (AM) has been used in Traditional Chinese Medicine (TCM) for about 2,000 years. In the study, we firstly determined the antioxidant levels of five AM extracts by •OH-scavenging, •O2−-scavenging, Fe2+-chelating, Cu2+-chelating, DPPH·-scavenging, and ABTS+·-scavenging assays. After measurement of the chemical contents in five AM extracts, we quantitatively analyzed the correlations between antioxidant levels and chemical contents. It was observed that total phenolics and total flavonoids had significant positive correlations with antioxidant levels (R = 0.685 and 0.479, respectively). In contrast, total sugars and total saponins presented lower correlations with antioxidant levels (R = −0.272 and 0.244, respectively). It means that antioxidant activity of AM should be attributed to total phenolics (including phenolic acids and flavonoids), and not total sugars and total saponins. Further analysis indicated that phenolic acids exhibited higher R values with radical-scavenging assays (R = 0.32–1.00), while flavonoids showed higher R values with metal-chelating assays (R= 0.86 and 0.90). In conclusion, AM exerts its antioxidant effect through metal-chelating, and radical-scavenging which is via donating hydrogen atom and donating electron. Its metal-chelating may result from flavonoids, while its radical-scavenging can be attributed to phenolic acids, especially caffeic acid, ferulic acid, and protocatechuic acid.

OBJECTIVE:To develop a HPLC-ELSD method for determination of ilexside II in leaves of Ilex cornuta.METHOD:The separation was performed on a Waters Syemmetry Shield RP18 column (4.6 mm x 250 mm, 5 microm) and methanol-water was used as mobile phase in gradient elution. The flow was 1.0 mL x min(-1), and the temperature was set at 35 degrees C.RESULT:The calibration curve showed good linearity in the test range (R2 = 0.9997). And the mean recovery was 101.3%, RSD was 2.0% (n=6).CONCLUSION:The developed HPLC-ELSD method was accurate and reproducible, and can be used for the determination of ilexside II in leaves of Ilex cornuta.