Cytotoxicity and underlying mechanism of evodiamine in HepG2 cells / 北京大学学报(医学版)

OBJECTIVE@#To investigate evodiamine (EVO)-induced hepatotoxicity and the underlying mechanism.@*METHODS@#HepG2 cells were treated with EVO (0.04-25 μmol/L) for different time intervals, and the cell survival rate was examined by cell counting kit-8 (CCK-8) method. After HepG2 cells were treated with EVO (0.2, 1 and 5 μmol/L) for 48 h, the alanine transaminase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), alkaline phosphatase (ALP) activities and total bilirubin (TBIL) content of supernatant were detected. A multifunctional microplate reader was used to detect the intracellular superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in HepG2 cells to evaluate the level of cell lipid peroxidation damage. The interactions between EVO and apoptosis, autophagy or ferroptosis-associated proteins were simulated by molecular docking. The HepG2 cells were stained by mitochondrial membrane potential (MMP) fluorescent probe (JC-10) and annexin V-fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI), and MMP and apoptosis in HepG2 cells were detected by flow cytometry. The protein expression levels of caspase-9, caspase-3, bile salt export pump (BSEP) and multidrug resistance-associated protein 2 (MRP2) were detected by Western blot.@*RESULTS@#The cell survival rate was significantly reduced after the HepG2 cells were exposed to EVO (0.04-25 μmol/L) in a time- and dose-dependent manner. The half maximal inhibitory concentration (IC50) of the HepG2 cells treated with EVO for 24, 48 and 72 h were 85.3, 6.6 and 4.7 μmol/L, respectively. After exposure to EVO (0.2, 1 and 5 μmol/L) for 48 h, the ALT, AST, LDH, ALP activities and TBIL content in the HepG2 cell culture supernatant, and the MDA content in the cells were increased, and SOD enzyme activity was decreased. Molecular docking results showed that EVO interacted with apoptosis-associated proteins (caspase-9 and caspase-3) better. JC-10 and Annexin V-FITC/PI staining assays demonstrated that EVO could decrease MMP and promote apoptosis in the HepG2 cells. Western blot results indicated that the protein expressions of cleaved caspase-9 and cleaved caspase-3 were upregulated in the HepG2 cell treated with EVO for 48 h. In contrast, the protein expressions of pro-caspase-3, BSEP and MRP2 were downregulated.@*CONCLUSION@#These results suggested that 0.2, 1 and 5 μmol/L EVO had the potential hepatotoxicity, and the possible mechanism involved lipid peroxidation damage, cell apoptosis, and cholestasis.

Exploratory screening of potential pan-cancer biomarkers based on The Cancer Genome Atlas database / 北京大学学报(医学版)

OBJECTIVE@#To screen potential pan-cancer biomarkers based on The Cancer Genome Atlas (TCGA) database, and to provide help for the diagnosis and prognosis assessment of a variety of cancers.@*METHODS@#"GDC Data Transfer Tool" and "GDCRNATools" packages were used to obtain TCGA database. After data sorting, a total of 13 cancers were selected for further analysis. False disco-very rate (FDR) < 0.05 and fold change (FC) >1.5 were used as the differential expression criteria to screen genes and miRNAs that were up- or down-regulated in all the 13 cancers. In the receiver operating characteristic curve (ROC curve), the area under the curve (AUC), the best cut-off value and the corresponding sensitivity and specificity were used to reflect diagnostic significance. The Kaplan-Meier method was used to calculate the survival probability and then the log-rank test was performed. Hazard ratio (HR) was calculated to reflect prognostic evaluation significance. DAVID tool were used to perform GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analysis for differentially expressed genes. STRING and TargetScan tools were used to analyze the regulatory network of differentially expressed genes and miRNAs.@*RESULTS@#A total of 48 genes and 2 miRNAs were differentially expressed in all the 13 cancers. Among them, 25 genes were up-regulated, 23 genes and 2 miRNAs were down-regulated. Most differentially expressed genes and miRNAs had good ability to distinguish between the cases and controls, with AUC, sensitivity and specificity up to 0.8-0.9. Survival analysis results show that differentially expressed genes and miRNAs were significantly associated with patient survival in a variety of cancers. Most up-regulated genes were risk factors for patient survival (HR>1), while most down-regulated genes were protective factors for patient survival (0 < HR < 1). The enrichment analysis of GO and KEGG showed that the differentially expressed genes were mostly enriched in biological events related to cell proliferation. In the regulatory network analysis, a total of 13 differentially expressed genes and 2 differentially expressed miRNAs had regulatory and interaction relationships.@*CONCLUSION@#The 48 genes and 2 miRNAs that were differentially expressed in 13 cancers may serve as potential pan-cancer biomarkers, providing help for the diagnosis and prognosis evaluation of a variety of cancers, and providing clues for the development of broad-spectrum tumor therapeutic targets.

EGFRvⅢ and immunotherapy of glioblastoma multiforme / 中国肿瘤生物治疗杂志

@#[Abstract] EGFRvIII (epidermal growth factor receptor variant Ⅲ) is the most common mutant of epidermal growth factor receptor, which expresses in over 30% glioblastoma multiforme (GBM). EGFRvIII results from deletion of exon 2-7, leading to its constitutive activation, which is closely related to tumorigenesis, development and poor prognosis of GBM. The unique glycine site on EGFRvIII provides ideal molecular target for immunotherapy, which possess great potential value of killing GBM cell, inhibiting progress and invasion. Encouraging progress has been achieved in peptide/DC vaccine, therapeutic antibody, small molecular inhibitor and adoptive immunotherapy experimentally and clinically. The characteristics of EGFRvIII and the development in its oriented immunotherapy were summarized in this review.

Diagnosis Performance of 99mTc-MIBI and Multimodality Imaging for Hyperparathyroidism / 华中科技大学学报（医学）（英德文版）

This study aimed to examine the diagnosis performance of 99mTc-methoxyisobutylisonitrisonitrile (99mTc-MIBI) and multimodality imaging [ultrasound,single-photon emission computed tomography/computed tomography (SPECT/CT)] for hyperparathyroidism (HPT).From Nov.2009 to Dec.2015,clinical data of a total of 43 HPT patients (16 males and 27 females;26-70 years old,average age:51.60±10.66 years old) were retrospectively analyzed.Among them,19 patients with primary hyperparathyroidism (PHPT) underwent 99mTc-MIBI planar imaging,24 [15 with PHPT and 9 with secondary hyperparathyroidism (SHPT)] underwent SPECT/CT hybrid imaging,and 41 (33 with PHPT and 8 with SHPT) had neck ultrasound imaging.Final diagnosis was determined by pathological examination after surgery.The positive rate was compared between different imaging modalities,and the correlation analysis was conducted between imaging results and lesion size or serum parathyroid hormone (PTH) level.The results showed that the total positive rates of 99mTc-MIBI imaging,ultrasound,and the two combined imaging in the 43 HPT cases were 90.70％ (39/43),58.54％ (24/41),and 100％ (41/41),respectively.According to lesion numbers,the positive rates were 79.10％ (53/67),53.23％ (33/62),and 88.71％ (55/62),respectively.SPECT/CT hybrid images were positive in all the 24 patients who underwent this examination.The mean maximum diameters of the lesions in 99mTc-MIBI positive and negative patients were 1.96±0.95 cm and 1.36±0.67 cm respectively,with statistically significant difference noted (P=0.03).The T/NT of 99mTc-MIBI imaging at the early phase was correlated positively with serum PTH level (r=0.40,P=0.01).The T/NT of 99mTc-MIBI imaging at both the early phase and the delay phase was correlated positively with lesion size (r=0.51,and r=0.45,respectively;P＜0.01 for both).It was concluded that 99mTc-MIBI imaging presents significant value for location diagnosis of HPT,especially when combined with SPECT/CT hybrid imaging or ultrasound.The 99mTc-MIBI uptake correlates positively with serum PTH level and lesion size.

Prediction of ETA oligopeptides antagonists from Glycine max based on in silico proteolysis / 中国中药杂志

Oligopeptides are one of the the key pharmaceutical effective constituents of traditional Chinese medicine(TCM). Systematic study on composition and efficacy of TCM oligopeptides is essential for the analysis of material basis and mechanism of TCM. In this study, the potential anti-hypertensive oligopeptides from Glycine max and their endothelin receptor A (ETA) antagonistic activity were discovered and predicted based on in silico technologies.Main protein sequences of G. max were collected and oligopeptides were obtained using in silico gastrointestinal tract proteolysis. Then, the pharmacophore of ETA antagonistic peptides was constructed and included one hydrophobic feature, one ionizable negative feature, one ring aromatic feature and five excluded volumes. Meanwhile, three-dimensional structure of ETA was developed by homology modeling methods for further docking studies. According to docking analysis and consensus score, the key amino acid of GLN165 was identified for ETA antagonistic activity. And 27 oligopeptides from G. max were predicted as the potential ETA antagonists by pharmacophore and docking studies.In silico proteolysis could be used to analyze the protein sequences from TCM. According to combination of in silico proteolysis and molecular simulation, the biological activities of oligopeptides could be predicted rapidly based on the known TCM protein sequence. It might provide the methodology basis for rapidly and efficiently implementing the mechanism analysis of TCM oligopeptides.

Chemical constituents of cortex periplocae radicis and its main toxic ingredients: Research advances / 国际药学研究杂志

Cortex Periplocae Radicis (CPR) is the dried root barks of Periploca spium Bge. It has been used in China for treat­ing rheumatoid arthritis and strengthening the bone and the musculature for thousands of years. Recently, an increasing number of bio­activities of CPR have been recognized, including tumor suppression and anti-chronic heart failure function. However, long-term use or large doses of CPR may produce adverse reactions, which constrains its applications in clinical therapy. Therefore, it is critical to know the chemical components of CPR in order to understand the toxicity mechanism. Nearly a hundred chemical compositions have been found, however, various classes, obfuscated names of different compounds, and disaccord between chemical structure and name were major obstacles to studying pharmacodynamics and toxicity of CPR. In this paper, 94 chemical components of CPR are classified and reviewed, which is valuable for the comprehensive understanding of its biological functions and safe clinical use.

Discovery of potential ATP-sensitive potassium channel openers with potential hypotensive activity from Chinese herbs based on molecular simulation / 中国中药杂志

In this research, a combined method of ligand-based pharmacophore (LBP), structure-based pharmacophore (SBP), and molecular docking was applied for virtual screening potential ATP-sensitive potassium channel (KATP) openers from Chinese herbs. LBP models were generated by 3D-QSAR pharmacophore(hypogen) program, based on the training set composed of 48 KATP agonists. The best LBP model consisted of one hydrogen-bond acceptor, one hydrogen-bond donor, one hydrophobic feature, one aromatic ring and five excluded volumes. Besides, the correlation coefficient of training set and test set, N, and CAI value of the model were 0.876 4, 0.705 8, 3.304, and 2.616 respectively. Meanwhile, SBP models were also generated based on a 3D structure of KATP (PMID: PM0079770). The best SBP model consisted of six hydrogen-bond acceptors, eight hydrogen-bond donors, seven hydrophobic features and eighteen excluded volumes. The corresponding N and CAI value were 2.200 and 2.017. Then, the best LBP model and SBP model were applied to identify potential KATP openers from Traditional Chinese Medicine Database(TCMD), respectively. 349 hits were obtained after analyzed by drug-likeness rules. Moreover, 12 compounds with high docking scores were reserved after molecular docking evaluation. Interestingly, part of the results had been verified as hypotensive active ingredients by literatures. Therefore, this study uncovers a specific target effect contained in TCMD, and provides candidates for new KATP openers' research.

Screen potential CYP450 2E1 inhibitors from Chinese herbal medicine based on support vector regression and molecular docking method / 中国中药杂志

Inhibition of cytochrome P450 (CYP450) enzymes is the most common reasons for drug interactions, so the study on early prediction of CYPs inhibitors can help to decrease the incidence of adverse reactions caused by drug interactions.CYP450 2E1(CYP2E1), as a key role in drug metabolism process, has broad spectrum of drug metabolism substrate. In this study, 32 CYP2E1 inhibitors were collected for the construction of support vector regression (SVR) model. The test set data were used to verify CYP2E1 quantitative models and obtain the optimal prediction model of CYP2E1 inhibitor. Meanwhile, one molecular docking program, CDOCKER, was utilized to analyze the interaction pattern between positive compounds and active pocket to establish the optimal screening model of CYP2E1 inhibitors.SVR model and molecular docking prediction model were combined to screen traditional Chinese medicine database (TCMD), which could improve the calculation efficiency and prediction accuracy. 6 376 traditional Chinese medicine (TCM) compounds predicted by SVR model were obtained, and in further verification by using molecular docking model, 247 TCM compounds with potential inhibitory activities against CYP2E1 were finally retained. Some of them have been verified by experiments. The results demonstrated that this study could provide guidance for the virtual screening of CYP450 inhibitors and the prediction of CYPs-mediated DDIs, and also provide references for clinical rational drug use.

Study on anti-hyperlipidemia mechanism of high frequency herb pairs by molecular docking method / 中国中药杂志

Traditional Chinese medicine (TCM) has definitely clinical effect in treating hyperlipidemia, but the action mechanism still need to be explored. Based on consulting Chinese Pharmacopoeia (2010), all the lipid-lowering Chinese patent medicines were analyzed by associated rules data mining method to explore high frequency herb pairs. The top three couplet medicines with high support degree were Puerariae Lobatae Radix-Crataegi Fructus, Salviae Miltiorrhizae Radix et Rhizoma-Crataegi Fructus, and Polygoni Multiflori Radix-Crataegi Fructus. The 20 main ingredients were selected from the herb pairs and docked with 3 key hyperlipidemia targets, namely 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase), peroxisome proliferator activated receptor-α (PPAR-α ) and niemann-pick C1 like 1 (NPC1L1) to further discuss the molecular mechanism of the high frequency herb pairs, by using the docking program, LibDock. To construct evaluation rules for the ingredients of herb pairs, the root-mean-square deviation (RMSD) value between computed and initial complexes was first calculated to validate the fitness of LibDock models. Then, the key residues were also confirmed by analyzing the interactions of those 3 proteins and corresponding marketed drugs. The docking results showed that hyperin, puerarin, salvianolic acid A and polydatin can interact with two targets, and the other five compounds may be potent for at least one of the three targets. In this study, the multi-target effect of high frequency herb pairs for lipid-lowering was discussed on the molecular level, which can help further researching new multi-target anti-hyperlipidemia drug.

Discovering L-type calcium channels inhibitors of antihypertensive drugs based on drug repositioning / 中国中药杂志

This study was amid to construct the pharmacophore model of L-type calcium channel antagonist in the application of screening Drugbank and TCMD. This paper repositions the approved drugs resulting from virtual screening and discusses the relocation-based drug discovery methods, screening antihypertensive drugs with L-type calcium channel function from TCMD. Qualitative hypotheses wre generated by HipHop separately on the basis of 12 compounds with antagonistic action on L-type calcium channel expressed in rabbit cardiac muscle. Datebase searching method was used to evaluate the generated hypotheses. The optimum hypothesis was used to search Drugbank and TCMD. This paper repositions the approved drugs and evaluates the antihypertensive effect of the chemical constituent of traditional Chinese medicine resulting from virtual screening by the matching score and literature. The results showed that optimum qualitative hypothesis is with six features, which were two hydrogen-bond acceptors, four hydrophobic groups, and the CAI value of 2.78. Screening Drugbank achieves 93 approved drugs. Screening TCMD achieves 285 chemical constituents of traditional Chinese medicine. It was concluded that the hypothesis is reliable and can be used to screen datebase. The approved drugs resulting from virtual screening, such as pravastatin, are potentially L-type calcium channels inhibitors. The chemical constituents of traditional Chinese medicine, such as Arctigenin III and Arctigenin are potentially antihypertensive drugs. It indicates that Drug Repositioning based on hypothesis is possible.

Virtual screening for natural CETP inhibitors by structure-based pharmacophore / 中国中药杂志

Cholesterol ester transfer protein (CETP) is a key regulator of high density lipoprotein (HDL). Owing to its important role in the reverse of cholesterol transport, CETP has become a hotspot target in modulating lipid drug design. In this paper, structure based pharmacophore (SBP) models for CETP inhibitors were built based on the protein structure 4F2A from Protein Database (PDB). The best pharmacophore contained six hydrophobic features, one hydrogen bond acceptor feature and nine excluded volume features, with the N and CAI value was 3.33 and 2.31 respectively. Then the model was used to search the traditional Chinese medicine database (TCMD) and 629 compounds originated from 315 TCM herbs were obtained. Molecular docking was also used to validate SBP by analyzing the critical amino acid residue and the interaction between potential active compounds and receptor. In this study, several TCM herbs, like Lycii Frutus and Schisandrae chinensis fructus, which contained more optimal SBP based screening results, have been reported hypolipidemic effect, and need to be studied deeply in a more focused research on herbal active constituents. Therefore, this study could provide reliable fundamental data for exploring the action mechanisms of TCM, and be applicable to identify lead candidates, which can be utilized as starting scaffolds for natural CETP inhibitors.

Application of support vector machine approach in studying nephron toxicity of Chinese medicinal materials / 中国中药杂志

On the basis of web databases, 111 compounds with nephrotoxicity and 90 compounds without nephrotoxicity were collected as data set of nephrotoxicity discrimination model, 39 compounds with tubular necrosis and 39 compounds without tubular necrosis were collected as data set of tubular necrosis discrimination model. The 6 122 molecular descriptors, including physicochemical, charge distribution and geometrical descriptors were calculated to characterize the molecular structure of the above-mentioned compounds. CfsSubsetEval valuation method and BestFirst-D1-N5 searching method were used to select molecular descriptors. Two models with high accuracy were built based on the support vector machine (SVM) approach, respectively. Accuracy, sensitivity, specificity and matthew's correlation coefficient of the two models were all above 70%. By using 22 nephrotoxicity compounds of Chinese medicine, the nephrotoxicity discrimination model was further verified with an accuracy of 72.73%. Using the tubular necrosis discrimination model, 10 potential compounds which can cause tubular necrosis were screened from the positive results of nephrotoxicity discrimination model, 6 of them have been verified by literatures. The results demonstrated that the discrimination models can be applied to screen nephrotoxic compounds from Chinese medicinal materials, and they also offer a new research idea for the further studies on the mechanism of nephrotoxicity.

Discovery of potential nicotinic acid receptor agonists from Chinese herbal medicines based on molecular simulation / 中国中药杂志

Nicotinic acid could increase high density lipoprotein and reduce serum total cholesterol, low density lipoprotein cholesterol and triglycerides in human bodies, thus is frequently applied in treating low high-density lipoprotein cholesterol and hypertriglyceridemia in clinic. However, according to the findings, nicotinic acid could also cause adverse effects, such as skin flush, beside its curative effects. In this study, bioisosterism, fragment-based search and Lipinski's Rule of Five were used to preliminarily screen out potential TCM ingredients that may have similar pharmacological effects with nicotinic acid from Traditional Chinese medicine database (TCMD). Afterwards, homology modeling and flexible docking were used to further screen out potential nicotinic acid receptor agonists. As a result, eleven candidate compounds were derived from eight commonly used traditional Chinese medicines. Specifically, all of the candidate compounds' interaction with nicotinic acid receptor was similar to nicotinic acid, and their docking scores were all higher than that of nicotinic acid, but their druggability remained to be further studied. Some of the eight source traditional Chinese medicines were used to lower lipid according to literature studies, implying that they may show effect through above means. In summary, this study provides basis and reference for extracting new nicotinic acid receptor agonists from traditional Chinese medicines and improving the medication status of hyperlipidemia.

Application of support vector machine in screening neurotoxic compounds from traditional Chinese medicine / 中国中药杂志

In this study, based on web database, 324 neurotoxic compounds and 234 non-neurotoxic compounds were selected as a data set for neurotoxicity discriminative model. 6 122 molecular descriptors, including charge distribution, physicochemical and geometrical descriptors,were calculated to characterize the molecular structure of neurotoxic compounds. The combination of Cfs Subset Evaluation and Best First-D1-N5 searching was used to select molecular descriptors. A discrimination model with high accuracy was built based on the support vector machine (SVM) approach. Meanwhile, the model accuracy, sensitivity and specificity were all above 80%. Besides, 30 traditional Chinese medicine compositions with neurotoxicity were set as external validation to further verify the model accuracy,with anaccuracy of 73.333%. Using the model, 13 potential neurotoxic compounds were screened from Sophorae subprostrate Radix,4 of them were verified by literatures. The results demonstrated that the discrimination model can be applied to screen neurotoxic compounds from Chinese medicinal materials.

Effect of ginsenoside Rb1 on central oxidative stress of rats with postoperative fatigue syndrome / 中草药

Objective: To investigate the central oxidative stress characteristics of rats with postoperative fatigue syndrome (POFS) and the antifatigue mechanism of ginsenosides Rb1. Methods: Rat models of POFS were established by using the 70% middle part of small bowel resection method. Ninety-six SD rats were randomly divided into control, model, and ginsenoside Rb1 (GRb1, 10 mg/kg) groups by weight. Rats in each group were administered 1 h before operation and were then divided into four subgroups at days 1, 3, 7, and 10. Morris water-maze test was done on postoperative days 2-7. Meanwhile, grasping test, malondialdehyde (MDA) content, superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) activities were detected on postperative days 1, 3, 7, and 10 and the ultrastructure of hippocampal CA1 area was observed through electron microscope. Results: Compared with the control group, the maximum grip of model rats had an obvious decline on days 3, 7, and 10 (P < 0.05). The total average escape latency was significantly extended (P < 0.05) and the platform crossing times were significantly reduced (P < 0.05). Compared with the model group, the above indexes of rats in GRb1 group were effectively improved after the intervention (P < 0.05). Compared with the control group, on postoperative days 1 and 3, the MDA content was obviously increased (P < 0.05) and SOD activity was obviously raised (P < 0.05). On postoperative day 7, GSH-Px activity was obviously raised (P < 0.05). After the intervention of GRb1, the MDA content was effectively decreased (P < 0.05), SOD and GSH-Px activities were effectively improved (P < 0.05). Electron microscope showed that the ultrastructure of hippocampal CA1 area of rats in GRb1 group was significantly improved. Conclusion: Surgical stress leads to the state change of central oxidative stress; GRb1 could reduce the damage of oxidative stress by strengthening the activity of central anti-oxidant enzymes, so as to protecte central neurons, which may be one of the mechanisms against POFS.

Investigation relationship between chronic benzene poisoning and ABO blood type / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the relevance between chronic benzene poisoning and ABO blood type.</p><p><b>METHODS</b>1014 benzene-exposed workers chosen from Shanghai and 4196 non-benzene-exposed workers chosen from Yangpu district were accepted the ABO blood type identification, and two groups of workers compared with the Han population of Shanghai and China for the ABO blood type distribution; the 71 cases of chronic benzene poisoning were compared with the group of benzene-exposed workers for the ABO blood type distribution.</p><p><b>RESULTS</b>There were no significant differences of the ABO blood type distribution among the Han population of China, the Han population of Shanghai and the group of non-benzene-exposed workers (x(2)=7.95, P>0.05). The A type blood distribution frequency in the group of chronic benzene poisoning patients was 42.25%, significantly higher than the group of benzene-exposed workers (29.48%), and there was statistically significant difference (x(2)=5.11,P<0.05). The B type blood distribution frequency in the group of chronic benzene poisoning patients was 12.68% , significantly lower than the group of benzene-exposed workers (25.15%), and there was statistically significant difference (x(2)=5.61, P<O.05). There were no significant differences of the O or AB type blood distribution frequency between the group of chronic benzene poisoning patients and the group of benzene-exposed workers (P>0.05).</p><p><b>CONCLUSION</b>The people with A type blood are susceptible to chronic benzene poisoning, however, the people with B type blood are not susceptible to chronic benzene poisoning.</p>